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# TITLE ABSTRACT
INTRODUCTION: SD Dengue Duo (Standard Diagnosis) commercial kit is an immunochromatographic rapid test that
detects NS1 protein and IgG/IgM dengue antibodies simultaneously., OBJECTIVE: to evaluate the operational and
functional characteristics of this system for the detection of virological and serological markers., METHODS: sera
panel was made up by 161 samples, 113 from patients with clinically and serologically confirmed dengue caused by
[Evaluation of the SD any of the four dengue virus serotypes and 48 negative samples. All these samples were tested by SD Dengue Duo Kit
Dengue Duo diagnosis and by Platelia Dengue NSI Ag, IgM Capture ELISA and ELISA Inhibition Method used as reference assays., RESULTS:
1. system for detection of the evaluated kit showed a 57.75% sensitivity for the detection of NS1 protein, false negatives were detected in
NS1 protein and IgM and samples collected 5 days or more after fever onset in secondary infection cases. IgM detection showed 96.0%
IgG dengue antibodies]. sensitivity and 98.4% specificity. Furthermore, high agreement (95.7%) in classifying dengue infection types (primary
or secondary infections) was observed. The global study of the 3 markers, the sensitivity rose to 100%.,
CONCLUSIONS: SD Dengue Duo is a simple, easy and rapid assay; it does not require additional equipment, can be
used for acute and convalescence serum samples and offers a good alternative for dengue diagnosis in those
laboratories where a complete dengue virus diagnosis is difficult to perform.
Currently, dengue fever is the most important re-emerging mosquito-borne viral disease, with the major proportion
of the target population residing in the developing countries of the world. In endemic areas, potentially fatal
secondary dengue infections, characterized by high anti-dengue IgG antibody titers, are most common. Most
currently available commercial dengue diagnostic kits rely on the use of whole virus antigens and are consequently
associated with false positives due to serologic cross-reactivity, high cost of antigen production, and biohazard risk.
This has prompted the need to develop an alternate antigen to replace the whole virus antigen in diagnostic tests. We
A custom-designed
have designed and expressed a novel recombinant protein antigen by assembling key immunodominant linear IgG-
recombinant
specific dengue virus epitopes, chosen on the basis of pepscan analysis, phage display, and computer predictions. The
2. multiepitope protein as a
recombinant dengue multiepitope protein was expressed to high levels in Escherichia coli, purified in a single step,
dengue diagnostic
yielding >25 mg pure protein per liter culture. We developed an in-house enzyme-linked immunosorbent assay
reagent.
(ELISA) to detect anti-dengue antibodies in a panel of 20 patient sera using the purified recombinant dengue
multiepitope protein as the capture antigen. The ELISA results were in excellent agreement with those obtained
using a commercially available diagnostic test, Dengue Duo rapid strip test from PanBio, Australia. The high epitope
density, careful choice of epitopes, and the use of E. coli system for expression, coupled to simple purification, jointly
have the potential to lead to the development of an inexpensive diagnostic test with a high degree of sensitivity and
specificity.
Dengue is a RNA viral illness of the genus Flavivirus which can cause, depending on the pervasiveness of the infection,
hemorrhagic dengue fever or dengue shock syndrome. Herein we present an electrochemical label free approach
A dual marker label free
enabling the rapid sensitive quantification of NS1 and IgG (supporting an ability to distinguish primary and secondary
electrochemical assay for
3. infections). Using a bifunctional SAM containing PEG moieties and a tethered redox thiol, both markers are detectable
Flavivirus dengue
across clinically relevant levels by label free impedance derived redox capacitance. A subsequent frequency specific
diagnosis
immittance function approach enables assaying (within seconds) with no impairment of analytical quality (linearity,
sensitivity and variance). © 2017 Elsevier B.V.
4. A rapid point-of-care test Dengue fever is caused by the dengue virus (DENV), and DENV1 is the prevalent epidemic serotype in south China. A
 for dengue virus-1 based new lateral flow assay (LFA) based on a near-infrared (NIR) fluorescent dye was developed to detect anti-DENV1 IgG
on a lateral flow assay antibodies. DyLight-800 was used as the marker conjugated to goat anti-human IgG antibodies, and recombinant
with a near-infrared dengue type 1 envelope protein was used as the capture protein on the test line. Twenty samples from patients
fluorescent dye. infected with DENV1 and 160 negative controls were analyzed using this new NIR-LFA. The results of the NIR-LFA
were compared with the results of Panbio Dengue IgG ELISA and the Dengue Duo IgM/IgG Cassette. Nineteen
confirmed DENV1-positive samples were identified by NIR-LFA, giving 95% (19/20) sensitivity. No significant
differences existed in the results when the 20 primary clinical samples were analyzed using NIR-LFA, Panbio ELISA,
and the Dengue Duo Cassette. However, NIR-LFA had a lower limit of detection than IgG ELISA and Duo IgM/IgG
Cassette did when analyzing a 2-fold dilution series of the 19 samples positively identified by NIR-LFA. When
incorporated with an NIR POCT device, the new NIR-LFA was rapid, easy to use, and highly sensitive in detecting
DENV1, and has potential for application to clinical diagnosis.Copyright © 2018 Elsevier B.V. All rights reserved.
An indirect enzyme-linked immunosorbent assay (ELISA) was developed to detect and differentiate the antibody
Antibody to the
responses to Japanese encephalitis (JE) virus nonstructural protein NS1 between infected and vaccinated individuals.
nonstructural protein
The results showed that all convalescent sera from JE patients contained NS1-specific IgG antibodies, while 65 and
NS1 of Japanese
40% of these sera showed detectable NS1-specific IgM and IgA antibodies, respectively. Specificity analysis showed
encephalitis virus:
that NS1-specific IgM and IgA antibodies from JE patients do not cross-react to dengue virus NS1 glycoprotein, while
5. Potential application of
IgG antibodies from 10% of JE patients showed significant cross-reaction to dengue virus NS1 glycoprotein. To
mAb-based indirect
differentiate infection from vaccination, the immune sera from 24 children vaccinated with inactivated JE vaccine
ELISA to differentiate
were analyzed. The data showed that none of these immune sera had detectable NS1-specific IgG antibodies. The
infection from
results demonstrated the potential application of JE NS1-specific indirect ELISA to differentiate infection from
vaccination
vaccination. © 2001 Elsevier Science Ltd.
Dengue is the most important disease caused by an arbovirus worldwide. Its clinical manifestations are very large
from asymptomatic infections to severe diseases with fatal outcome. No effective antiviral treatment or vaccine is
available. Thus, a rapid and accurate diagnosis is of paramount importance both for better clinical case management
and surveillance. Diagnosis methods depend on the time clinical signs appeared.Within the 7 first days of fever, direct
Biological diagnosis of tests are preferred. RT-PCR methods are sensitive, specific, and can identify viral serotypes. Conventional RT-PCR will
6.
dengue probably be replaced by real time PCR as soon as standardised and accurate assays for the four serotypes will be
available. Serology (EIA) is used only after 7 days of disease, i.e. late in the course of dengue; it is accurate, specific
but not discriminatory for serotypes and high cross-reactive. NS1 antigen detection still lack of clinical sensitivity and
viral isolation is too fastidious. Even though ameliorations are necessary, viral detection by RT-PCR remains the best
tool in clinical settings for a rapid diagnosis of severe dengue infections.
Clinical and virological Background: Detection of dengue NS1 antigen in acute infection has been proposed for early diagnosis of dengue
factors influencing the disease. The aim of this study was to evaluate the clinical and virological factors influencing the performance of the
performance of a ns1 Platelia NS1 Ag kit (BioRad) and to assess the potential use of NS1 antigen and dengue viral loads as markers of
7. antigen-capture assay dengue disease severity. Methodology/Principal Findings: Blood specimens were collected from patients hospitalized
and potential use as a at the Kampong Cham hospital during the 2006 and 2007 dengue epidemics in Cambodia. Dengue infection was
marker of dengue confirmed in 243/339 symptomatic patients and in 17 asymptomatic individuals out of 214 household members
disease severity tested. Overall sensitivity and specificity of Platelia NS1 Ag kit were 57.5% and 100% respectively. NS1 Ag assay
 combined with IgM antibody capture ELISA significantly increased the sensitivity for dengue diagnosis. NS1 Ag
positivity rate was found significantly higher in DF than in DHF/DSS, in primary than in secondary infections, in
patients with a high viremia (>5 log/mL) and in patients infected with DENV-1. In asymptomatic individuals, the NS1
Ag capture sensitivity tends to be lower than that in symptomatic patients. Milder disease severity was observed
independently in patients with RNA copy number >5 log10 cDNA equivalents/mL or in high level of NS1 antigen ratio
or in DENV-1 infection. Conclusions: Overall sensitivity of NS1 Ag detection kit varied widely across the various forms
of dengue infection or disease. Sensitivity was highest in patients sampled during the first 3 days after onset of fever,
in patients with primary infection, DENV-1 infection, with high level of viremia and in DF rather than DHF/DSS. In
asymptomatic patients, RT-PCR assay has proved to be more sensitive than NS1 antigen detection. The NS1 antigen
level correlated significantly with viremia and a low NS1 antigen ratio was associated with more severe disease. ©
2011 Duong et al.
In 2014, a large outbreak of dengue occurred in Guangzhou, China. This outbreak prompted us to evaluate NS1 and
RNA for the early diagnosis of acute dengue infection, in addition to the combination with IgM antibody. We aimed to
find the differences of three assays about dengue diagnosis. This study was an evaluation of diagnosis test. Based on
WHO criteria 2009, dengue RNA, NS1, and IgM/IgG were detected from 294 patients (180 dengue patients, 114 non-
Clinical evaluation of dengue patients) by three diagnostic kits made in China. The χ2 test, sensitivity, and specificity were used in
dengue RNA, NS1, and statistical analysis. The ratios of dengue patients with low platelet counts (<100×109/L 32.2%) or white blood cell
8. IgM for diagnosis of counts (<4.0×109/L 58.9%) were significantly higher compared to non-dengue patients (P<0.05). Dengue NS1
dengue in Southern was shown sensitive (93.9%) for diagnostic use. RNA had a better performance with 98.1% of sensitivity from day 1 to
China day 4 after illness onset. IgM performed better at day 5 or more with 74.0% of sensitivity. The diagnostic rate using a
combination of RNA and IgM was 97.8% and 96.7% using NS1 and IgM. A patient with low platelet and white blood
cell counts needs additional tests for dengue during an epidemic. RNA and NS1 were most valuable for early diagnosis
of dengue, whereas IgM was best suited as a supplementary method for patients at day 5 or more after illness onset.
© 2015 Wiley Periodicals, Inc.
Background & objectives: Rapid diagnostic test (RDT) kits are widely used in India for the diagnosis of dengue
infection. It is important to evaluate the validity and reliability of these RDTs. The study was aimed to determine the
sensitivity, specificity and predictive value of four commercially available RDTs [Panbio Dengue Duo cassette,
Comparative evaluation
Standard Diagnostics (SD) Bioline Dengue Duo, J. Mitra Dengue Day-1 test and Reckon Dengue IgG/IgM] against
of validity and cost-
composite reference criteria (CRC), and compare the cost of the tests. Methods: In this prospective observational
benefit analysis of rapid
study for diagnostic accuracy, we tested stored blood samples from 132 cases of dengue and 149 controls of other
diagnostic test (RDT) kits
infections as classified based on CRC, with all the four RDTs. The CRC was based on the epidemiological
9. in diagnosis of dengue
considerations, common clinical features and laboratory abnormalities. The non-dengue controls were the cases of
infection using
proven alternative diagnosis. The diagnostic performances of the tests were compared in terms of sensitivity,
composite reference
specificity and predictive value along with the cost involved per test. Results: The sensitivity of the Panbio and SD RDT
criteria: A cross-sectional
kits was found to be 97.7 and 64.3% respectively, and the specificities were 87.8 and 96.6% respectively. The
study from south India
sensitivity of the NS1 antigen capture by SD Duo, Reckon, J. Mitra RDTs was 20.9, 18.6 and 27.1% respectively. The
prevalence of dengue specific IgG antibody with Panbio RDT kits was 49.3%. The cost per test for Panbio, SD, Reckon
and J. Mitra is US$ 6.90, 4.27, 3.29 and 3.61 respectively. Conclusion: It was concluded that in dengue outbreak,
 Panbio IgM capture RDT alone is reliable and easily available test which can be used in acute phase of dengue
infection in any resource limited set up. NS1 capture rates by any of the other three RDTs might not be reliable for the
diagnosis of acute dengue infection. © 2016, Malaria Research Center. All rights reserved.
Dengue fever (DF) is endemic in India and dengue hemorrhagic fever (DHF) has been reported with increasing
frequency in the last decade. We evaluated three commercial assays for detection of antibodies to dengue virus, to
assess their performance in a diagnostic laboratory. Sera from 58 patients collected during a febrile outbreak in New
Comparative evaluation
Delhi in 1997 were studied. The methods evaluated were MRL Diagnostic Dengue Fever Virus IgM Capture ELISA, Pan
of various commercial
10. Bio Dengue Duo IgM and IgG Capture ELISA and Pan Bio Rapid Immunochromatographic test. The MRL ELISA correctly
assays for diagnosis of
identified 97.8% (43 of 44) of samples as dengue positive while the Pan Bio Duo ELISA and Pan Bio RIT identified
dengue fever
95.45% (42 of 44). The sensitivities of both Pan Bio Duo ELISA and Pan Bio RIT for primary dengue and secondary
dengue were 100% and 93.54% respectively. The specificity of three assays were MRL IgM ELISA 100%, Pan Bio Duo
ELISA 92.8% and Pan Bio RIT 85.7%.
Difficulties in the management of dengue infections include the lack of rapid diagnostic methods and symptoms of
dengue are often confused with those of other diseases. Five commercial kits were evaluated for the detection of
Comparison of five dengue-specific IgM and IgG antibodies using sera obtained from patients with primary and secondary dengue
serological diagnostic infections, as well as other febrile illnesses. All kits were compared to the in-house IgM ELISA and HI assays. The rapid
assays for the detection test kits took either 30 to 45 min (PanBio Dengue Duo Cassette, Accusens Dengue Virus Rapid Strip Test, and Unitest
11.
of IgM and IgG Dengue IgM and IgG Combo Rapid Test) or between 2 to 4 h (PanBio Dengue Duo Capture ELISA, and Antivirus IgM
antibodies to dengue Detecion Kit 96 [Pentax Corporation]). Most kits were able to detect IgM in more than 90% of the secondary
virus convalescent sera, while IgG detection was generally high (80 to 100%). All five kits showed high specificity when
tested against sera from other febrile patients, and have been shown to be extremely useful in the diagnosis of
dengue infections.
Eighty sera from patients with dengue virus infections, 24 from patients with Japanese encephalitis (JE) (Japanese
encephalitis virus infection), and 78 from patients with nonflavivirus infections, such as malaria (Plasmodium
Comparison of PanBio
infection), typhoid (Salmonella typhi infection), leptospirosis (Leptospira infection), and scrub typhus (Orientia
Dengue Duo enzyme-
tsutsugamushi infection), were used. The MRL test showed superior sensitivity for dengue virus infections (94 vs.
linked immunosorbent
89%), while the PanBio test showed superior specificity for JE (79 vs. 25%) and other infections (100 vs. 91%). The
assay (ELISA) and MRL
PanBio ELISA showed better overall performance, as assessed by the sum of sensitivity and specificity (F value). When
dengue fever virus
12. dengue virus and nonflavivirus infections were compared, F values of 189 and 185 were obtained for the PanBio and
immunoglobulin M
MRL tests, respectively, while when dengue virus infections and JE were compared, F values of 168 and 119 were
capture ELISA for
obtained. The results obtained with individual sera in the PanBio and MRL IgM ELISAs showed good correlation, but
diagnosis of dengue virus
this analysis revealed that the cutoff value of the MRL test was set well below that of the PanBio test. Comparing the
infections in Southeast
sensitivity and specificity of the tests at different cutoff values (receiver-operator analysis) revealed that the MRL and
Asia.
PanBio IgM ELISAs performed similarly in distinguishing dengue virus from nonflavivirus infections, although the
PanBio IgM ELISA showed significantly better distinction between dengue virus infections and JE.
Comparison of PanBio The performances of the MRL dengue fever virus immunoglobulin M (IgM) capture enzyme-linked immunosorbent
13. dengue duo enzyme- assay (ELISA) and the PanBio Dengue Duo IgM capture and IgG capture ELISA were compared. Eighty sera from
linked immunosorbent patients with dengue virus infections, 24 sera from patients with Japanese encephalitis (JE), and 78 sera from
 assay (ELISA) and MRL patients with nonflavivirus infections, such as malaria, typhoid, leptospirosis, and scrub typhus, were used. The MRL
dengue fever virus test showed superior sensitivity for dengue virus infections (94 versus 89%), while the PanBio test showed superior
immunoglobulin M specificity for JE (79 versus 25%) and other infections (100 versus 91%). The PanBio ELISA showed better overall
capture ELISA for performance, as assessed by the sum of sensitivity and specificity (F value). When dengue virus and nonflavivirus
diagnosis of dengue virus infections were compared, F values of 189 and 185 were obtained for the PanBio and MRL tests, respectively, while
infections in Southeast when dengue virus infections and JE were compared, F values of 168 and 119 were obtained. The results obtained
Asia. with individual sera in the PanBio and MRL IgM ELISAs showed good correlation, but this analysis revealed that the
cutoff value of the MRL test was set well below that of the PanBio test. Comparing the sensitivity and specificity of
the tests at different cutoff values (receiver-operator analysis) revealed that the MRL and PanBio IgM ELISAs
performed similarly in distinguishing dengue virus from nonflavivirus infections, although the PanBio IgM ELISA
showed significantly better distinction between dengue virus infections and JE. The implications of these findings for
the laboratory diagnosis of dengue are discussed.
Background: A number of commercial ELISA for dengue diagnosis have recently become available, though direct
comparison between these assays have not been published. Objectives: The Venture Technologies Dengue IgM and
IgG Dot Blot assays and the PanBio Dengue Duo IgM and IgG Capture ELISA were compared. Study design: Paired sera
Comparison of PanBio from patients with dengue (n=20) and Japanese encephalitis (JE, n=10), and single sera from patients with typhoid
Dengue Duo Igm and IgG (n=10), leptospirosis (n=10) and scrub typhus (n=10) were assayed according to the manufacturer's instructions.
Capture ELISA and Results: The Dot Blot IgM ELISA showed higher sensitivity than the PanBio IgM ELISA (100 vs. 95%), while the PanBio
14.
Venture Technologies IgM ELISA showed higher specificity in JE (100 vs. 20%) and non-flavivirus infections (100 vs. 97%). Defining elevation
Dengue IgM and IgG Dot of either IgM or IgG as a positive result, the Dot Blot and ELISA tests both showed 100% sensitivity in dengue
Blot infection, while the PanBio test showed superior specificity in JE (70 vs. 0%) and non-flavivirus infections (100 vs.
67%). Conclusions: Both assays are useful aids to the serological diagnosis of dengue infection. The clinical setting,
user preference and local conditions will be important in determining which test is more appropriate. Copyright (C)
2000 Elsevier Science B.V.
The Venture Technologies Dengue IgM and IgG Dot Blot assays and the PanBio Dengue Duo IgM and IgG Capture
ELISA were compared. Paired sera from patients with dengue virus (n=20) from Thailand and Japanese encephalitis
Comparison of PanBio virus (JE, n=10) from Vietnam, and single sera from patients with typhoid (Salmonella typhi infection) (n=10) from
Dengue Duo IgM and IgG Malaysia, leptospirosis (n=10) from Australia and scrub typhus (Orientia tsutsugamushi infection) (n=10) from
Capture ELISA and Thailand were assayed according to the manufacturer's instructions. The Dot Blot IgM ELISA showed higher sensitivity
15.
Venture Technologies than the PanBio IgM ELISA (100 vs. 95%), while the PanBio IgM ELISA showed higher specificity in JE (100 vs. 20%) and
Dengue IgM and IgG Dot non-flavivirus infections (100 vs. 97%). Defining elevation of either IgM or IgG as a positive result, the Dot Blot and
Blot. ELISA tests both showed 100% sensitivity in dengue infection, while the PanBio test showed superior specificity in JE
(70 vs. 0%) and non-flavivirus infections (100 vs. 67%). It is concluded that both assays are useful aids to the
serological diagnosis of dengue infection.
Comparison of seven Seven commercial assays were evaluated to determine their suitability for the diagnosis of acute dengue infection: (i)
commercial antigen and the Panbio dengue virus Pan-E NS1 early enzyme-linked immunosorbent assay (ELISA), second generation (Alere,
16.
antibody enzyme-linked Australia); (ii) the Panbio dengue virus IgM capture ELISA (Alere, Australia); (iii) the Panbio dengue virus IgG capture
immunosorbent assays ELISA (Alere, Australia); (iv) the Standard Diagnostics dengue virus NS1 antigen ELISA (Standard Diagnostics, South
 for detection of acute
dengue infection
Comparison of the
diagnostic accuracy of
17. commercial NS1-based
diagnostic tests for early
dengue infection
Comparison of two
dengue NS1 rapid tests
for sensitivity, specificity
18.
and relationship to
viraemia and antibody
responses
Korea); (v) the Standard Diagnostics dengue virus IgM ELISA (Standard Diagnostics, South Korea); (vi) the Standard
Diagnostics dengue virus IgG ELISA (Standard Diagnostics, South Korea); and (vii) the Platelia NS1 antigen ELISA (Bio-
Rad, France). Samples from 239 Thai patients confirmed to be dengue virus positive and 98 Sri Lankan patients
negative for dengue virus infection were tested. The sensitivities and specificities of the NS1 antigen ELISAs ranged
from 45 to 57% and 93 to 100% and those of the IgM antibody ELISAs ranged from 85 to 89% and 88 to 100%,
respectively. Combining the NS1 antigen and IgM antibody results from the Standard Diagnostics ELISAs gave the best
compromise between sensitivity and specificity (87 and 96%, respectively), as well as providing the best sensitivity for
patients presenting at different times after fever onset. The Panbio IgG capture ELISA correctly classified 67% of
secondary dengue infection cases. This study provides strong evidence of the value of combining dengue virus
antigen- and antibody-based test results in the ELISA format for the diagnosis of acute dengue infection. Copyright
© 2012, American Society for Microbiology.
Background: We compared the diagnostic accuracy and reproducibility of commercially available NS1-based dengue
tests and explored factors influencing their sensitivities. Methods: Paired analysis of 310 samples previously
characterized as positive (n = 218) and negative (n = 92) for viral isolation and/or RT-PCR and/or IgM seroconversion.
Masked samples were tested by two observers with Platelia (TM) Dengue NS1 Ag, second generation Pan-E (TM)
Dengue Early ELISA, SD Dengue NS1 Ag ELISA, Dengue NS1 Ag STRIP (TM), and SD BIOLINE (TM) Dengue Duo
(NS1/IgM/IgG). Results: SD BIOLINE (TM) NS1/IgM/IgG had the highest sensitivity (80.7% 95% CI 75-85.7) with
likelihood ratios of 7.4 (95% CI 4.1-13.8) and 0.21 (95% CI 0.16-0.28). The ELISA-format tests showed comparable
sensitivities; all below 75%. STRIP (TM) and SD NS1 had even lower sensitivities (<65%). The sensitivities significantly
decreased in samples taken after 3 days of fever onset, in secondary infections, viral serotypes 2 and 4, and severe
dengue. Adding IgM or IgG to SD NS1 increased its sensitivity in all these situations. Conclusions: The simultaneous
detection of NS1/IgM/IgG would be potentially useful for dengue diagnosis in both endemic and non endemic areas.
A negative result does not rule out dengue. Further studies are required to assess the performance and impact of
early laboratory diagnosis of dengue in the routine clinical setting.
Background: Dengue is a major public health problem in tropical and subtropical countries. Rapid and easy diagnosis
of dengue can assist patient triage and care-management. The detection of DENV NS1 on rapid lateral flow tests
offers a fast route to a presumptive dengue diagnosis but careful evaluations are urgently needed as more and more
people use them.Methods: The sensitivity and specificity of the Bio-Rad NS1 Ag Strip and SD Dengue Duo
(NS1/IgM/IgG) lateral flow rapid tests were evaluated in a panel of plasma samples from 245 Vietnamese patients
with RT-PCR confirmed dengue and 47 with other febrile illnesses.Results: The NS1 rapid tests had similar diagnostic
sensitivities (respectively 61.6% and 62.4%) in confirmed dengue cases but were 100% specific. When IgM/IgG results
from the SD Dengue Duo were included in the test interpretation, the sensitivity improved significantly from 62.4%
with NS1 alone to 75.5% when NS1 and/or IgM was positive and 83.7% when NS1 and/or IgM and/or IgG was
positive. Both NS1 assays were significantly more sensitive for primary than secondary dengue. NS1 positivity was
associated with the underlying viraemia as NS1-positive samples had a significantly higher viraemia than NS1-negative
samples.Conclusions: These data suggest that the NS1 test component of these assays are highly specific and have
similar levels of sensitivity. The IgM parameter in the SD Duo test improved overall test sensitivity without
compromising specificity. The SD Dengue Duo lateral flow rapid test deserves further prospective evaluation in
 dengue endemic settings. © 2010 Tricou et al; licensee BioMed Central Ltd.
Dengue diagnosis was one of the topics discussed at the symposium 'The Global Threat of Dengue - Desperately
Seeking Solutions' organized during the 10th International Congress of Infectious Diseases held in Singapore in 2002.
In this paper, a review is presented focusing on the main advances, problems and challenges of dengue diagnosis. IgM
capture ELISA, virus isolation in mosquito cell lines and live mosquitoes, dengue specific monoclonal antibodies and
Dengue diagnosis, PCR have all represented major advances in dengue diagnosis. However, an appropriate rapid, early and accessible
19.
advances and challenges diagnostic method useful both for epidemiological surveillance and clinical diagnosis is still needed. Also, tools that
suggest a prognosis allowing for better management are also needed. Finally, laboratory infrastructure, technical
expertise and research capacity must be improved in endemic countries in order to positively influence dengue
surveillance, clinical case management and the development of new approaches to dengue control. © 2003
International Society for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.
Dengue is a major health problem in many parts of the tropical world presenting as both epidemic & endemic forms.
It is a mosquito borne illness caused by one of the serotypes of dengue viruses. Mortality rate of dengue in untreated
patients is 10%-12%. The present study aims to determine whether the rapid test SD dengue duo (IgG, IgM, and NS-1
Ag detection) can be used as screening test in dengue outbreak when compared to ELISA. 226 serum samples were
Dengue Duo Rapid Test:
tested in patients clinically suspected Dengue. All the 226 samples were subjected to IgG, IgM Microlisa test. The
A Valuable Screening
20. same were put on rapid SD bioline Dengue duo rapid test and was compared with ELISA. 150 samples were tested
Test for Dengue
positive with ELISA (either positive for IgG, IgM). When compared with ELISA, Rapid test showed sensitivity of 80.6%
Outbreak
specificity and positive predictive value of 100% & zero false positive rates. Efficiency of the test was 87.16% SD
Dengue duo rapid test should be a valuable screening test for dengue fever outbreak which can be interpreted easily.
Results were comparable to ELISA. It provides additional diagnostic investigation of primary and secondary dengue
that compliments NS-1 antigen detection.
Rapid diagnosis of dengue is crucial for proper patient care. As IgM antibody appears early during the disease course,
its detection is a valuable tool for rapid diagnosis. We evaluated and compared two commercial tests, the PanBio
Rapid Immunochromatographic Card Test (Brisbane, Australia) and the PanBio Microwell IgM ELISA with an IgM
Capture ELISA (National Institute of Virology, Pune, India). A total of 154 samples from individuals with febrile illness
Dengue fever: Its having dengue fever (DF)-like symptoms were tested. The NIV IgM Capture ELISA (MAC-ELISA) showed a high
laboratory diagnosis, positivity rate (38.9%) as compared to the PanBio Rapid (22.7%) and the PanBio IgM ELISA (20.7%). The NIV MAC-
21.
with special emphasis on ELISA showed a high sensitivity (96%) as compared to PanBio Rapid (73%) and PanBio IgM ELISA (72%). But the
IgM detection specificity was low (81%) when compared to PanBio Rapid (95%) and PanBio IgM ELISA (97%) using the latent class
analysis model. The MAC-ELISA, though a three-day procedure, should be a valuable screening test because of its high
sensitivity rates. But rapidity gets compromised, as batch testing is required along with technical difficulty in
performance. The "Rapid" test is an easier option in small peripheral laboratories in India because of its obvious
advantages.
Dengue virus 4 (DENV-4) Dengue fever (DF) is a mosquito-borne viral disease of great concern in tropical and subtropical regions of the world.
re-emerges after 30 One important cause of the increase in DF is rapid development and urbanization has led to proliferation of the Aedes
22.
years in Brazil: aegypti mosquito, the vector responsible for transmission of the illness. Surveillance of dengue virus (DENV) infection
Cocirculation of DENV-2, in Brazil shows the predominance of DENV-1, DENV-2, and DENV-3 until 2010. This study reports the reappearance of
 DENV-3, and DENV-4 in DENV-4 in Brazil for the first time in 30 years. Serum samples were collected from individuals (n = 214) exhibiting
Bahia fever and muscular pain in Bahia, Brazil, during 2011–2012. These samples were subjected to reverse transcription-
polymerase chain reaction (RT-PCR)/nested PCR, which revealed that 82% of samples were positive for DENV-4; most
were older age groups and exhibited a serological pattern consistent with a primary infection. The cocirculation of
multiple DENV serotypes within the same city places the population at risk for a fatal form of the disease. Therefore,
with the increasing incidence of severe DF cases, early diagnosis will be a priority for public health efforts in Brazil. ©
2015, National Institute of Health. All rights reserved.
Saliva was collected prospectively from patients presenting with suspected dengue infection 4 to 8 days after the
onset of symptoms and assayed by a commercial dengue immunoglobulin M (IgM) and IgG capture enzyme- linked
immunosorbent assay (ELISA) (PanBio Dengue Duo ELISA). Laboratory diagnosis was based on virus isolation and on
Detection of specific hemagglutination inhibition (HAI) assay and an in-house IgM and IgG capture ELISA. With a positive result defined as
23. antibodies in saliva either salivary IgM or IgG levels above the cutoff value, an overall sensitivity of 92% was obtained for both primary-
during dengue infection and secondary- dengue patients (22 of 24), while no patients with non-flavivirus infections (n = 11) and no healthy
laboratory donors (n = 17) showed elevation of salivary antidengue antibody (100% specificity). Salivary IgG levels
correlated well with serum HAI titer (r = 0.78), and salivary IgG levels could be used to distinguish between primary-
and secondary-dengue virus infections.
WHO recommendations for dengue diagnosis require laboratory facilities. Antibody-based rapid diagnostic tests
(RDTs) have performed poorly, and clinical diagnosis remains the mainstay in dengue-endemic countries. We
evaluated a combination antigen-antibody RDT for point-of-care testing in a high-prevalence setting. In this
prospective cohort study, adults were enrolled from a tertiary infectious disease centre for evaluation of
undifferentiated febrile illness from October 2011 to May 2012. SD Bioline Dengue Duo was evaluated at point-of-
Diagnosing Dengue at care against a WHO-based reference standard of viral isolation, RT-PCR, NS1-, IgM-, and IgG-ELISA. 246 adults were
the Point-of-Care: Utility enrolled (median age 34 years, range 18-69), of which 197 could be confirmed definitively as either dengue or non-
24. of a Rapid Combined dengue. DENV-2 was the predominant serotype (79.5%) and the ratio of primary to secondary cases was 1:1.1. There
Diagnostic Kit in were no test failures and minimal interobserver variation with a Fleiss' kappa of 0.983 (95% CI 0.827-1.00). Overall
Singapore sensitivity and specificity were 93.9% (95% CI 88.8-96.8%) and 92.0% (95% CI 81.2-96.9%) respectively. Using WHO
clinical criteria alone for diagnosis had similar sensitivities (95.9%, 95% CI 91.4-98.1%) and lower specificities (20.0%,
95% CI 11.2-33.0%). No significant difference in performance was found when testing early versus late presenters,
primary versus secondary cases, or DENV-1 versus DENV-2 infections. The use of a combination RDT fulfills WHO
ASSURED criteria for point-of-care testing and can enhance dengue diagnosis in an endemic setting. This has the
potential to markedly improve clinical management of dengue in the field.
The Dengue IgM Capture ELISA (MAC-ELISA) is the immunoenzymatic system recommended by the Pan American
Diagnosis of Dengue
Health Organization and the World Health Organization for the serological diagnosis of dengue virus infection due to
Virus Infection by the
its high sensitivity, ease of performance, and use of a single acute-phase serum sample. However, tests with this
Visual and Simple
25. enzyme-linked immunosorbent assay (ELISA) system are time-consuming and require equipment for washing,
AuBioDOT
incubation, and reading of the results. AuBioDOT is a multistep visual diagnostic immunoassay that uses technology
Immunoglobulin M
based on the immunoglobulin M (IgM) capture ELISA principle. This system uses white polyethylene opaque plates as
Capture System
the solid phase, colloidal gold as the marker, and silver ion amplification. It does not require special equipment, it is
 totally manually operated, and it can be performed in less than 1 h. The sensitivity and specificity of AuBioDOT for the
detection of anti-dengue virus IgM antibodies were studied with a panel of 336 serum samples (150 serum samples
from patients with suspected or serologically confirmed dengue virus infection, 186 serum samples from healthy
blood donors and patients without dengue virus infection). The results were compared with those obtained by the
MAC-ELISA. A sensitivity of 97.7% and a specificity of 97.1% were obtained. The concordance of the two tests was
97.3%, with a kappa index of 0.94. The application of AuBioDOT for the detection of anti-dengue virus IgM antibodies
is recommended as an alternative method for the diagnosis of dengue virus infection, both for clinical diagnosis and
for seroepidemiological surveillance. The system is useful under field conditions and in laboratories and requires little
equipment.
Dengue is known for its serious life-threatening complications. New rapid kits available recently in India target
Early dengue diagnosis
circulating non-structural protein (NS1) antigen from day one onwards. The sensitivity and specificity of a newly
by nonstructural protein
introduced rapid combo kit against two conventional ELISA kits is assessed. The performance of this kit is quite
1 antigen detection:
satisfactory since excellent agreement of 94.26% was observed with particular reference to NS1 antigen detection
Rapid
26. among all three kits namely Rapid SD Bioline dengue Duo (SD Korea), InBios DENV Detect NS1 ELISA, USA and dengue
immunochromotography
Early ELISA, Panbio, Australia. The false positivity of the rapid kit is very low since its specificity as for as NS1 antigen
versus two the enzyme-
detection is concerned is 98.33%. The use of combination kit helps to detect additional cases of dengue, which are
linked immunosorbent
negative for NS1 antigen but positive for IgM and/or IgG antibodies, thus facilitating early diagnosis in remote areas
assay kits
and small laboratorie.
A commercial Dengue Duo rapid test kit was evaluated for early dengue diagnosis by detection of dengue virus NS1
antigen and immunoglobulin M (IgM)/IgG antibodies. A total of 420 patient serum samples were subjected to real-
Early diagnosis of time reverse transcription-polymerase chain reaction (RT-PCR), in-house IgM capture enzyme-linked immunosorbent
Dengue infection using a assay (ELISA), hemagglutination inhibition assay, and the SD Dengue Duo rapid test. Of the 320 dengue acute and
commercial Dengue Duo convalescent sera, dengue infection was detected by either serology or RT-PCR in 300 samples (93.75%), as compared
27.
rapid test kit for the with 289 samples (90.31%) in the combined SD Duo NS1/IgM. The NS1 detection rate is inversely proportional,
detection of NS1, IGM, whereas the IgM detection rate is directly proportional to the presence of IgG antibodies. The sensitivity and
and IGG. specificity in diagnosing acute dengue infection in the SD Duo NS1/IgM were 88.65% and 98.75%, respectively. The
assay is sensitive and highly specific. Detection of both NS1 and IgM by SD Duo gave comparable detection rate by
either serology or RT-PCR.
High levels of anti-dengue IgM or IgG can be detected using numerous rapid diagnostic tests (RDTs). However, the
sensitivity and specificity of these tests are reduced by changes in envelope glycoprotein antigenicity that inevitably
Enhanced performance occur in limited expression systems. A novel RDT was designed to enhance diagnostic sensitivity. Dengue viruses
of an innovative dengue cultured in animal cells were used as antigens to retain the native viral coat protein. Monoclonal antibodies (mAbs)
IgG/IgM rapid diagnostic were then developed, for the first time, against domain I of envelope glycoprotein (EDI). The anti-dengue EDI mAb
28.
test using an anti-dengue was employed as a capturer, and EDII and EDIII, which are mainly involved in the induction of neutralizing antibodies
EDI monoclonal antibody in patients, were fully available to bind to anti-dengue IgM or IgG in patients. A one-way automatic blood separation
and dengue virus antigen device prevented reverse migration of plasma and maximize the capture of anti-dengue antibodies at the test lines. A
clinical evaluation in the field proved that the novel RDT (sensitivities of 96.5% and 96.7% for anti-dengue IgM and
IgG) is more effective in detecting anti-dengue antibodies than two major commercial tests (sensitivities of 54.8% and
 82% for SD BIOLINE; 50.4% and 75.3% for PanBio). The innovative format of RDT can be applied to other infectious
viral diseases.
A commercially available capture ELISA for the detection of specific IgM and IgG antibodies produced during dengue
infection (PanBio Dengue Duo) was evaluated with paired serum specimens from 176 patients in Singapore. Diagnosis
Evaluation of a
was based on a haemagglutination inhibition (HAI) assay, with patients having either primary dengue (n = 90),
commercial capture
secondary dengue (n = 58) or no dengue (n = 28) infection. The combined use of IgM and IgG (sensitivity, 99%;
enzyme-linked
specificity, 96%) was superior to the use of IgM alone (sensitivity, 88%; specificity, 96%) or IgG alone (sensitivity, 85%;
immunosorbent assay
29. specificity, 96%). Furthermore, with the first serum sample of the pair of serum samples, the ELISA was able to
for detection of
diagnose significantly more cases of dengue than the HAI assay (55% vs 14%). The results of the IgG capture ELISA
immunoglobulin M and
gave a significant correlation with those of the HAI assay (r = 0.91; P<0.0001), and the IgG capture ELISA could be used
G antibodies produced
to distinguish between primary and secondary infection. The best distinction was observed when an IgG cutoff ratio
during dengue infection.
of 3.0 was used, with 88% of primary infections and 98% of secondary infections being correctly classified. This ELISA
should prove to be useful in the clinical diagnosis of dengue infection.
Early diagnosis is important for clinical management of dengue disease. While classic laboratory tests are often
tedious and time consuming, point of care devices offer a rapid, cost-effective and user-friendly alternative provided
their accuracy is acceptable. This study evaluated the sensitivity, specificity and efficiency of SD BIOLINE Dengue
Duo® rapid NS1, IgM and IgG test kit for diagnosis of acute dengue virus infection. Standard laboratory diagnostics,
EVALUATION OF A RT-PCR, IgM and IgG capture ELISAs were carried out on 143 suspected dengue patient samples obtained from a Sri
COMMERCIAL RAPID Lankan population. Using the results of these standard laboratory tests as reference, the sensitivity and specificity of
30. TEST KIT FOR DETECTION the SD Dengue Duo® NS1 test was 57% and 87%, respectively, and those of the IgM test was 50% and 84%,
OF ACUTE DENGUE respectively. The combined sensitivity and specificity of the SD Dengue Duo® NS1/ IgM test was 72% and 80%,
INFECTION respectively. The SD Dengue Duo® NS1 test detected NS1 for up to 9 days from onset of fever. Primary and
secondary dengue cases were classified according to the IgG test, of which the kit identified 88% and 26% of primary
and of secondary infection, respectively. Although the SD Dengue Duo® kit was not as accurate as the standard
tests, it still can serve the useful reference for initial screening of suspected dengue cases, especially in poor resource
hospital settings and aid in clinical disease management of dengue infection.
This study has evaluated the performance of a rapid immunochromatographic test (ICT) device in detecting
Evaluation of a rapid
antibodies to Dengue virus (DENV) in a tertiary hospital in South India. Sera from hospital attendees, with requests for
immunochromatographi
DENV antibody testing, were tested with the Panbio Dengue Duo Cassette and a reference antibody capture assay for
c device for the
the detection of IgM (Dengue IgM capture ELISA-National Institute of Virology, India) and IgG (Dengue IgG capture
detection of IgM & IgG
31. ELISA-Panbio Diagnostics Inc., Australia) antibodies. The ICT results were compared with results of antibody capture
antibodies to Dengue
tests for the detection of the IgM and IgG antibodies, respectively. Accuracy indices for IgM and IgG detection,
viruses (DENV) in a
respectively were- sensitivity 81.8% and 87.5%, specificity 75.0%, and 66.6%, positive predictive value (PPV) 61.0%
tertiary care hospital in
and 72.9% and negative predictive value (NPV) 89.6% and 83.9%. The device performs poorly in detection of IgM and
South India
IgG antibodies to DENVs and is not recommended for use as a stand-alone diagnostic test.
Evaluation of a rapid Serology is the mainstay of diagnosis in Dengue virus infection. Various rapid tests for antibody detection have been
32. immunochromatographi developed. They can prove to be important diagnostic tools especially in the field set up due to technical simplicity.
c test in the diagnosis of We evaluated a Rapid immunochromatographic assay. The rapid test was performed on acute phase sera collected
 dengue fever from patients suspected to be suffering from Denguefever/DHF. These samples were then tested by Dengue Duo
Capture ELISA and compared. The rapid test showed a good sensitivity for the detection of secondary dengue
infection and thus can be a good screening tool.
Various assays have been developed to diagnose dengue virus infection, relying on techniques from the fields of
serology and molecular biology. Many of these assays have been successful, but there is still an urgent need for
accurate, simple and rapid diagnostic assays to diagnose dengue virus infection and to assist in patient management.
Evaluation of an
Using a panel of well-characterized sera and a collection of retrospective samples obtained during the dengue
immunoglobulin M-
epidemics that occurred in Belém, Brazil, between 2002 and 2009, a modified immunoglobulin M-specific capture
specific capture enzyme-
33. enzyme-linked immunosorbent assay (Rapid-MAC-ELISA) was evaluated and compared with the "gold standard" MAC-
linked immunosorbent
ELISA, in order to assess the specificity, sensitivity, stability, reproducibility and cost-effectiveness of this new assay.
assay for rapid diagnosis
These results demonstrated that the Rapid-MAC-ELISA is comparable to the MAC-ELISA in terms of sensitivity and
of dengue infection
specificity and is highly reproducible; additionally, it is easily performed, less expensive than other available formats
and can be completed within three hours. Furthermore, the Rapid-MAC-ELISA can be used for the diagnosis of
dengue virus infections in resource-limited areas where dengue is endemic. © 2010 Elsevier B.V.
Background: Rapid diagnosis of dengue infection is essential to patient management and disease control. The
development of a rapid (5 min) immunochromatographic test and a 2 h commercial capture enzyme linked
immunosorbent assay (ELISA) for anti-dengue IgM and IgG antibodies may lead to more rapid and accurate testing in
Evaluation of capture peripheral health settings and diagnostic laboratories. Objectives: Evaluate two new commercial tests for dengue
ELISA and rapid serology (Dengue Rapid test and Dengue Duo ELISA; PanBio, Brisbane, Australia). Study design: The sensitivity and
immunochromatographi specificity of the tests were compared with in-house dengue IgM ELISA and hemagglutination-inhibition (HAI) assays
c test for the using known positive and negative dengue specimens, as well as specimens from non-dengue cases. Results: Both
34.
determination of IgM assays showed excellent sensitivity in the diagnosis of both primary and secondary dengue infection (100%). In both
and IgG antibodies assays, IgG levels showed excellent correlation with hemagglutination-inhibition (HAI) assay, and these could be used
produced during dengue to distinguish between primary and secondary dengue infections (92 and 97% of patients correctly classified in the
infection rapid test and Duo ELISA, respectively). Specificity in both assays was 89% when sera from patients, with no apparent
dengue infection, typhoid, leptospirosis and malaria, were tested. Conclusions: These tests should be a useful aid in
confirming the clinical diagnosis of dengue infection. The rapid test will be particularly valuable in peripheral health
settings, while the ELISA has a place in central testing laboratories.
Two new commercial tests for dengue serology, the Dengue Rapid test and Dengue Duo ELISA (PanBio, Brisbane,
Evaluation of capture Australia), were evaluated. The sensitivity and specificity of the tests were compared with in-house dengue IgM ELISA
ELISA and rapid and haemagglutination-inhibition (HAI) assays using known positive and negative dengue specimens, as well as
immunochromatographi specimens from non-dengue cases. Both assays showed excellent sensitivity in the diagnosis of both primary and
c test for the secondary dengue infection (100%). In both assays, IgG levels showed excellent correlation with the HAI assay, and
35.
determination of IgM these could be used to distinguish between primary and secondary dengue infections (92 and 97% of patients
and IgG antibodies correctly classified in the rapid test and Duo ELISA, respectively). Specificity in both assays was 89% when sera from
produced during dengue patients, with no apparent dengue infection, typhoid, leptospirosis and malaria, were tested. These tests should be a
infection. useful aid in confirming the clinical diagnosis of dengue infection. The rapid test will be particularly valuable in
peripheral health settings, while the ELISA has a place in central testing laboratories.
 During an active surveillance study in school children in Medellin, we assessed the performance of two diagnostic
strategies for dengue virus. A total of 41 patients with suspected dengue acute infection were evaluated. Diagnostic
strategies consisted of one combining Panbio (R) Dengue virus IgM and IgG Capture ELISAs (enzyme-linked
immunosorbent assays) with reverse transcriptase polymerase chain reaction (T-PCR) and another using a commercial
Evaluation of
rapid SD Bioline Dengue Duo (IgG/IgM + NS1 Ag) test. These two strategies were compared with the enzyme linked
Commercially Available
immunospot microneutralization test (ELISPOT-MNT). The sensitivity and specificity were 53.9% and 80.0% for the
Assays for Diagnosis of
combination of Panbio (R) ELISAs and RT-PCR tests, and 30.8% and 73.3% for the SD Bioline Duo test, respectively.
36. Acute Dengue in
ELISPOT-MNT detected 16.4% additional cases and revealed the presence of neutralizing antibodies in all the acute
Schoolchildren during an
samples, evidencing that they were all secondary infections. In contrast, Panbio (R) and SD Dengue Duo rapid tests
Epidemic Period in
only classified 23.0% and 26.9% of the cases as secondary dengue infections, respectively. Cohen's kappa coefficient
Medellin, Colombia
and McNemar's association tests demonstrated a significant disagreement between the two diagnostic strategies and
ELISPOT-MNT. Overall, these results evidence the relatively poor performances of commercial assays for the diagnosis
of acute and secondary dengue infections, compared with ELISPOT-MNT, and raise concern about the accuracy of
these assays for the diagnostic of dengue in endemic areas.
Evaluation of NS1Ag and
IgM antibodies against
37. dengue, importance for 0
epidemiological
surveillance
We determined the diagnostic performance of the OneStep NS1 and the OneStep IgG/IgM RDT kits against a panel of
samples which comprised of 174 dengue positive and 165 dengue negative sera characterized by three reference
Evaluation of OneStep
enzyme-linked immunosorbent assays (ELISAs). The diagnostic sensitivities of the OneStep kits for the detection of
Dengue NS1 RapiDipâ„¢
individual biomarkers of NS1, IgM and IgG were 90% (95% CI: 82.1–94.7), 32.4% (95% CI: 24.8–40.8) and 44.4%
InstaTest and OneStep
(95% CI: 38.2–50.7), respectively. The combination of the OneStep IgG/IgM kit with the OneStep NS1 kit
38. Dengue Fever IgG/IgM
demonstrated significantly higher sensitivities for the combined NS1/IgM (96.8%; 95% CI: 90.9–99.3) and
RapiCardâ„¢ InstaTest
NS1/IgM/IgG (99.5%; 95% CI: 97.1–99.9)(P < 0.001). In conclusion, the OneStep NS1 kit has high sensitivity and
during the course of a
specificity and is highly recommended for use. The low sensitivities for IgG (44.4%) and for IgM (32.4%) of the
dengue type 1 epidemic
OneStep IgG/IgM kit when used alone suggest it is best used in combination with the OneStep NS1 kit to enhance its
overall diagnostic performance. © 2017 Elsevier Inc.
Evaluation of six Six assays were evaluated in this study to determine their suitability for the diagnosis of acute dengue infection using
commercial point-of-care samples from 259 Sri Lankan patients with acute fevers (99 confirmed dengue cases and 160 patients with other
tests for diagnosis of confirmed acute febrile illnesses): (i) the Merlin dengue fever IgG & IgM combo device (Merlin), (ii) the Standard
acute dengue infections: Diagnostics Dengue Duo nonstructural 1 (NS1) antigen and IgG/IgM combo device (Standard Diagnostics, South
39. The need for combining Korea), (iii) the Biosynex Immunoquick dengue fever IgG and IgM (Biosynex, France) assay, (iv) the Bio-Rad NS1
NS1 antigen and IgM/IgG antigen strip (Bio-Rad, France), (v) the Panbio Dengue Duo IgG/IgM Cassette (Inverness, Australia), and (vi) the Panbio
antibody detection to dengue NS1 antigen strip (Inverness, Australia). The median number of days of fever prior to admission sample
achieve acceptable levels collection was 5 days (interquartile range, 3 to 7 days). Sensitivity and specificity of the NS1 antigen tests ranged from
of accuracy 49 to 59% and from 93 to 99%, respectively, and sensitivity and sensitivity of the IgM antibody test ranged from 71 to
 80% and from 46 to 90%, respectively. Combining the NS1 antigen and IgM antibody results from the Standard
Diagnostics Dengue Duo test gave the best compromise of sensitivity and specificity (93% and 89%, respectively) and
provided the best sensitivity in patients presenting at different times after fever onset. The Merlin IgM/IgG antibody
tests correctly classified 64% and 86% of the primary and secondary dengue infection cases, respectively, and the
Standard Diagnostics IgM/IgG antibody tests correctly classified 71% and 83% of the primary and secondary dengue
infection cases, respectively. This study provides strong evidence of the value of combining dengue antigen- and
antibody-based test results in the rapid diagnostic test (RDT) format for the acute diagnosis of dengue. Copyright ©
2011, American Society for Microbiology. All Rights Reserved.
The performance of six commercially available immunoassay systems for the detection of dengue virus-specific
immunoglobulin M (IgM) and IgG antibodies in serum was evaluated. These included two IgM and IgG enzyme
immunoassays (EIA) from MRL Laboratories and PanBio, a rapid immunochromatographic test (RIT) from PanBio,
immunofluorescence assays (IFA) from Progen, a dot blot assay from Genelabs, and a dipstick EIA from Integrated
Diagnostics (INDX). For this study a panel of 132 serum samples, including 90 serum samples from patients with
suspected dengue virus infection and 42 serum samples from patients with other viral infections, was used. In
addition, serial serum samples from two monkeys experimentally immunized and challenged with dengue virus type 2
Evaluation of six
were used. Results were considered conclusive when concordant results were obtained with four of the six antibody-
immunoassays for
specific assays. Based on this definition, the calculated overall agreement for the human serum samples for the
detection of dengue
40. respective IgM immunoassays was 97% (128 of 132), with 34% (45 of 132) positive serum samples, 63% (83 of 132)
virus-specific
negative samples, and 3% of samples (4 of 132) showing discordant results. The calculated overall agreement for the
immunoglobulin M and
IgG assays was 94% (124 of 132), with 49% (65 of 132) positive, 45% (59 of 132) negative, and 6% (8 of 132)
G antibodies
discordant results, respectively. The sensitivities of the dengue virus-specific assays evaluated varied between 71 and
100% for IgM and between 52 and 100% for IgG, with specificities of 86 to 96% and 81 to 100%, respectively. The
relative sensitivities of the respective IgM assays measured with the monkey serum samples were comparable with
those obtained with 12 serial serum samples from humans. Overall performance, based on the sum of the agreement,
sensitivity, specificity, and Kappa statistics of the IgM and IgG immunoassays, showed that the antibody detection
systems from INDX and Genelabs and the MRL and PanBio EIA are useful and reliable assays for dengue virus
serodiagnosis.
In this study, we evaluated the performance of a rapid test, the SD BIOLINE Dengue Duo (SD BDD) kit, with a panel of
serum samples from 310 Mexican patients with diagnosis of dengue infection previously confirmed by reference
Evaluation of the SD
enzyme-linked immunosorbent assay tests. Eighty-seven negative samples from other febrile illnesses were included
BIOLINE dengue duo
as controls. The SD BDD showed an overall sensitivity of 90.65% and specificity of 89.66%. No statistically significant
rapid test in the course
differences were found in the sensitivity of the SD BDD kit compared between primary or secondary infections
41. of acute and
(87.05% versus 93.57%, respectively, P = 0.0761) and dengue fever or dengue hemorrhagic fever cases (90.77% versus
convalescent dengue
89.74%, respectively, P = 0.7716). However, a higher sensitivity in the acute phase of dengue infection was found
infections in a Mexican
compared with the convalescent phase (93.03% versus 81.82%, respectively, P = 0.0089). These results indicate that
endemic region
the SD BDD kit is a useful tool to diagnose dengue infections, both in primary or secondary infections and mainly
during the acute phase. © 2014 Elsevier Inc.
42. Evaluation of the SD Introduction: SD Dengue Duo (Standard Diagnosis) commercial kit is an immunochromatographic rapid test that
 Dengue Duo diagnosis detects NS1 protein and IgG/IgM dengue antibodies simultaneously. Objective: To evaluate the operational and
system for detection of functional characteristics of this system for the detection of virological and serological markers. Methods: Sera panel
NS1 protein and IgM and was made up by 161 samples, 113 from patients with clinically and serologically confirmed dengue caused by any of
IgG dengue antibodies. the four dengue virus serotypes and 48 negative samples. All these samples were tested by SD Dengue Duo Kit and by
Platelia Dengue NS1 Ag, IgM Capture ELISA and ELISA Inhibition Method used as reference assays. Results: The
evaluated kit showed a 57.75% sensitivity for the detection of NS1 protein, false negatives were detected in samples
collected 5 days or more after fever onset in secondary infection cases. IgM detection showed 96.0% sensitivity and
98.4% specificity. Furthermore, high agreement (95.7%) in classifying dengue infection types (primary or secondary
infections) was observed. The global study of the 3 markers, the sensitivity rose to 100%. Conclusions: SD Dengue Duo
is a simple, easy and rapid assay; it does not require additional equipment, can be used for acute and convalescence
serum samples and offers a good alternative for dengue diagnosis in those laboratories where a complete dengue
virus diagnosis is difficult to perform.
: Dengue virus infection causes life-threatening hemorrhagic fever. Increasing evidence implies that dengue viral
Facilitation of Cell
nonstructural protein 1 (NS1) exhibits a tendency to elicit potentially hazardous autoantibodies, which show a wide
Adhesion by Immobilized
spectrum of specificity against extracellular matrix and platelet antigens. How NS1 elicits autoantibodies remains
Dengue Viral
unclear. To address the hypothesis that NS1 and matrix proteins may have structural and functional similarity, cell-
Nonstructural Protein 1
matrix and cell-NS1 interactions were evaluated using a cell-adhesion assay. The present study showed that dengue
43. (NS1): Arginine-Glycine-
NS1 immobilized on coverslips resulted in more cell adhesion than did the control proteins. This cell adhesion was
Aspartic Acid Structural
inhibited by peptides containing arginine-glycine-aspartic acid (RGD), a motif important for integrin-mediated cell
Mimicry within the
adhesion. In addition, anti-NS1 antibodies blocked RGD-mediated cell adhesion. Although there is no RGD motif in the
Dengue Viral NS1
NS1 protein sequence, these data indicate that RGD structural mimicry exists within the NS1 antigen, (C) Copyright
Antigen.
Oxford University Press 2002.
Background: Dengue diagnosis is complex and until recently only specialized laboratories were able to definitively
confirm dengue infection. Rapid tests are now available commercially making biological diagnosis possible in the field.
The aim of this study was to evaluate a combined dengue rapid test for the detection of NS1 and IgM/IgG antibodies.
The evaluation was made prospectively in the field conditions and included the study of the impact of its use as a
point-of-care test for case management as well as retrospectively against a panel of well-characterized samples in a
Field Evaluation and reference laboratory. Methodology/Principal Findings: During the prospective study, 157 patients hospitalized for a
Impact on Clinical suspicion of dengue were enrolled. In the hospital laboratories, the overall sensitivity, specificity, PPV and NPV of the
Management of a Rapid NS1/IgM/IgG combination tests were 85.7%, 83.9%, 95.6% and 59.1% respectively, whereas they were 94,4%, 90.0%,
44.
Diagnostic Kit That 97.5% and 77.1% respectively in the national reference laboratory at Institut Pasteur in Cambodia. These results
Detects Dengue NS1, IgM demonstrate that optimal performances require adequate training and quality assurance. The retrospective study
and IgG showed that the sensitivity of the combined kit did not vary significantly between the serotypes and was not affected
by the immune status or by the interval of time between onset of fever and sample collection. The analysis of the
medical records indicates that the physicians did not take into consideration the results obtained with the rapid test
including for care management and use of antibiotic therapy. Conclusions: In the context of our prospective field
study, we demonstrated that if the SD Bioline Dengue Duo kit is correctly used, a positive result highly suggests a
dengue case but a negative result doesn't rule out a dengue infection. Nevertheless, Cambodian pediatricians in their
 daily practice relied on their clinical diagnosis and thus the false negative results obtained did not directly impact on
the clinical management. © 2012 Andries et al.
Background: Dengue virus (DENV) infection is an emerging arboviral infection in tropical and sub-tropical countries in
South-East Asia, the Western Pacific and South and Central America. Secondary DENV infection is the most widely
accepted risk factor for the development of severe forms. Methods to discriminate primary and secondary DENV
infection may be of great prognostic value. ELISA based detection of specific antibodies (IgG and IgM) to the four
DENV serotypes is valuable for detemination of primary or secondary infection. Several studies had been performed
to discriminate primary and secondary DENV infection using the ratio of IgG and IgM at the various days of symptoms
onset. The aim of this study is to determine the best cut-off point of IgG to IgM ratio is able to discriminating
Immunoglobulin G (IgG) secondary from primary DENV infection in adult using samples from early days of symptoms onset. Methods: This
to IgM ratio in secondary prospective cohort study on 48 adult patients with DENV infected patients on the period of August 2011-January 2012
45. adult dengue infection in 5 out-patient settings health facilities in Tangerang district, Banten province, Indonesia with chief complaint of
using samples from early fever less than 3 days. Datas were collected on the day the patients attended health facilities, consisted of
days of symptoms onset demographic, clinical, laboratory, and serological data. Serological data from 48 serum sample from adult patients
were evaluated using Focus Diagnostics Dengue Virus IgM and IgG Capture DxSelect (TM) ELISA Kits to determine IgG,
IgM index values and SD Bioline Dengue Duo (TM) Rapid Tests to determine NS1, IgG, and IgM result. Results:
According to NS1, IgG and IgM results, 36 patients were classified as secondary infection, 12 were primary Infection.
The mean (SD) of IgG/IgM ratios for secondary and primary infection were 3.28 (0.54) and 0.18 (0.11) consecutively.
The IgG/IgM ratio of >= 1.14 confirmed secondary infection with sensitivity of 80.56 %, specificity 91.67 %, accuracy
level 83.33 %, and likely hood ratio of (LR) 9.67. Conclusion: The IgG/IgM ratio of >= 1.14 as the best cut off point to
determine secondary DENV infection in early days of symptoms onset.
Dengue is an arthropod borne disease that has become important worldwide. There is still no specific drug available
for treatment and also no protective vaccine that can be used. As such, specific diagnosis is essential to enable good
management and prevention of large outbreaks. Diagnosis today in many countries is still based on serology though
the detection of NS1 has slowly become incorporated. Diagnosis is critical for early intervention with specific
Laboratory diagnosis of preventive health measures to prevent fatalities and also to curtail spread and reduce economic losses. Serological
46.
dengue: A review assays mainly detect IgM which now as a single test is invalid unless a second sample is taken to confirm. As such to
effectively diagnose dengue at all stages of infection, assays with two or more markers are required or two samples
taken a few days apart. Other commonly used tests include NS1 detection, nucleic acid amplification and IgG
detection. However the sensitivities of the current commercial kits vary quite considerably and have to be interpreted
with caution. Hence knowledge of this disease is essential when conducting diagnostics for dengue.
The dengue duo rapid strip test (DDRST) was evaluated with 37 travellers admitted for febrile syndrome at the
Low specificity of an
University Hospitals of Marseilles, France, with blood smear positive for Plasmodium falciparum. The patients were
immunochromatographi
hospitalized between October 2001 and April 2002 after returning from the Comoros islands (n=21), Madagascar
c serological assay for
47. (n=1), West Africa (n=9), East Africa (n=1), India (n=1), French Polynesia (n=2) and West Indies (n=2). Eighteen sera
diagnosis of dengue
tested positive for the presence of immunoglobulin (Ig)M. Of these, only two were confirmed by ELISA (54.3%
fever in travelers
specificity). All 19 sera testing negative for IgM with DDRST were found negative with ELISA (100% negative predictive
returning with malaria.
value). Positive predictive value was 11.1%. The patients who exhibited a false-positive IgM detection by DDRST were
 returning from Comoros Islands (n=13), East Africa (n=2) and West Indies (n=1). It is concluded that the elevated rate
of false-positive IgM detected by DDRST is certainly not an immunological cross-reactivity due to poor specificity of
the recombinant envelope protein.
This paper describes a model for predicting hemoglobin (Hb) by using bioelectrical impedance analysis (BIA) in dengue
patients in the Hospital Universiti Kebangsaan Malaysia (HUKM). Bioelectrical impedance measurements were
Modeling of hemoglobin conducted on 83 (47 males and 36 females) serologically confirmed dengue fever (DF) and dengue hemorrhagic fever
in dengue fever and (DHF) patients during their hospitalization. The predictive equation for Hb was derived using multivariate analysis. We
48. dengue hemorrhagic investigated all the parameters in BIA, patients' symptom and demographic data. In this developed model, four
fever using bioelectrical predictors (reactance (Xc), sex, weight and vomiting) were found to be the best predictive factors for modeling Hb in
impedance dengue patients. However, the model can only explain approximately 42% of the variation in Hb status, thus single
frequency bio-impedance stand-alone technique is insufficient to monitor Hb for the DF and DHF patients. Further
investigation using multi-frequency BIA is recommended in modeling Hb to achieve the most parsimonious model.
We evaluated four dengue diagnostic devices from Alere, including the SD Bioline Dengue Duo (nonstructural [NS] 1
Ag and IgG/IgM), the Panbio Dengue Duo Cassette (IgM/IgG) rapid diagnostic tests (RDTs), and the Panbio dengue
IgM and IgG capture enzyme―linked immunosorbent assays (ELISAs) in a prospective, controlled, multicenter study
in Peru, Venezuela, Cambodia, and the United States, using samples from 1,021 febrile individuals. Archived,
well―characterized samples from an additional 135 febrile individuals from Thailand were also used. Reference
Multicountry prospective
testing was performed on all samples using an algorithm involving virus isolation, in―house IgM and IgG capture
clinical evaluation of two
ELISAs, and plaque reduction neutralization tests (PRNT) to determine the infection status of the individual. The
enzyme-linked
primary endpoints were the clinical sensitivities and specificities of these devices. The SD Bioline Dengue Duo had an
49. immunosorbent assays
overall sensitivity of 87.3% (95% confidence interval [CI], 84.1 to 90.2%) and specificity of 86.8% (95% CI, 83.9 to
and two rapid diagnostic
89.3%) during the first 14 days post―symptom onset (p.s.o.). The Panbio Dengue Duo Cassette demonstrated a
tests for diagnosing
sensitivity of 92.1% (87.8 to 95.2%) and specificity of 62.2% (54.5 to 69.5%) during days 4 to 14 p.s.o. The Panbio IgM
dengue fever
capture ELISA had a sensitivity of 87.6% (82.7 to 91.4%) and specificity of 88.1% (82.2 to 92.6%) during days 4 to 14
p.s.o. Finally, the Panbio IgG capture ELISA had a sensitivity of 69.6% (62.1 to 76.4%) and a specificity of 88.4% (82.6
to 92.8%) during days 4 to 14 p.s.o. for identification of secondary dengue infections. This multicountry prospective
study resulted in reliable real―world performance data that will facilitate data―driven laboratory test choices for
managing patient care during dengue outbreaks.
Objectives: Most of the serological methods, traditional and recent, that have been developed to assess laboratory
confirmation of dengue infection, suffer from the need to use separate tests for IgM and for IgG and the need for
paired serum specimens in order to establish a clinical interpretation. Moreover, most of the IgG tests ignore early
New dengue antibody convalescence as well as past exposure. The objectives of this study is to demonstrate the advantages of the
assay with unique ImmunoComb Dengue BiSpot, a new dengue antibody assay, that detects simultaneously both dengue-specific IgM
50.
differential detection of and IgG, in determining type and status of infection in a single test. Designs and methods: Single and paired positive
IgG and IgM antibodies and negative specimens obtained from various dengue-high prevalent regions and from dengue-free areas were used.
The specimens were comprised of admission samples of suspected patients and secondary dengue infection
individuals, a group of hospital patients, asymptomatic to dengue who originated from dengue high prevalence areas
and dengue-negative volunteers. All samples were tested by the ImmunoComb Dengue BiSpot kit along with the gold
 standard assays of Hemagglutination-inhibition (HAI) and MAC ELISA. Results: Testing 365 gold standard positive
specimens from different regions, the ImmunoComb Dengue BiSpot kit revealed sensitivity for IgM of 97.5% and
99.1% for IgG. The IgG was segregated into 2 groups: 86.6% for high titer and 12.5% for low titer of IgG. The overall
sensitivity was 98.9%. The overall specificity of the kit, testing 657 samples, mostly from dengue-free area was 97.9%.
The higher diagnostic resolution of the Dengue BiSpot test was demonstrated when the performance of the kit on the
various groups was compared to the gold standard tests and the dengue status was determined in paired specimens
by the new assay. Conclusions: The results of the present studies demonstrate the advantages of ImmunoComb
Dengue BiSpot assay as an alternate strategy for the determination of dengue infection status. The combined use of
IgM and IgG to discriminate between primary and secondary infections, on one hand, and providing high resolution in
distinction of convalescence stage and past infection, on the other hand, endow the kit with improved capacity in
assessing the clinical status of a tested individual in a single test. © 2008 The Canadian Society of Clinical Chemists.
Objectives: Non-structural protein 1 (NS1)-based tests may offer a larger window of opportunity for dengue diagnosis
and could constitute a very useful diagnostic tool. The aim of this study was to establish the overall accuracy of NS1-
based tests for diagnosing dengue infection. Methods: A meta-analysis was conducted including 18 studies published
up to October 1, 2012 identified using PubMed, ISI Web of Science, Google Scholar, and the Chinese National
Knowledge Infrastructure (CNKI) database. Results: For the single NS1-based tests - ELISA (Panbio Dengue Early ELISA
Kit, Dengue NS1 Ag ELISA Kit, and Platelia Dengue NS1 Ag-ELISA Kit) and immunochromatography (Dengue NS1 Ag
NS1-based tests with
STRIP Kit and SD BIOLINE Dengue Duo Strip Kit) - the summarized sensitivities and specificities were 67% (95%
diagnostic utility for
confidence interval (CI) 59-74%) and 99% (95% CI 97-99%), and 71% (95% CI 61-79%) and 99% (95% CI 98-100%),
51. confirming dengue
respectively. The hierarchical summary receiver operating characteristics (HSROCs) were 0.92 and 0.96, respectively.
infection: A meta-
For NS1 combined with an anti-dengue-specific IgM test, the summarized sensitivity, specificity, and HSROC were 83%
analysis
(95% CI 68-92%), 86% (95% CI 79-91%), and 0.91 (95% CI 0.89-0.93), respectively. The accuracy for serotypes was
50.0-90.9% for DENV-1, 38.5-85.7% for DENV-2, 46.7-91.3% for DENV-3, and 21.7-87.0% for DENV-4. Conclusions:
These results support the use of single NS1-based tests; they have good diagnostic utility for confirming dengue and
for distinguishing serotypes DENV-1 and 3 from DENV-2 and 4, while they can be used as a screening tool when
combined with an IgM test. Moreover, the Dengue NS1 Ag STRIP Kit appears to be the best for confirming and
serotyping dengue infection. © 2014 The Authors.
Dengue virus is endemic in French Guiana with occurrence of cyclical outbreaks. There is a need for rapid tests
allowing dengue laboratory diagnosis in healthcare centers scattered throughout this wide Amazonian territory. Our
objective was to evaluate the real-life performance of the SD BIOLINE Dengue Duo (IgG/IgMÂ +Â NS1 Ag) rapid test
(RDT) during the 2012–2013 dengue epidemics. The RDT was evaluated in parallel with routine laboratory tests,
Prospective evaluation of
PlateliaTM Dengue NS1 Ag and Focus Diagnostics Dengue Fever Virus IgM Capture DxSelect. A total of 3,347 patients
the SD BIOLINE Dengue
52. with suspected dengue acute infection were evaluated. The diagnostic performances of the SD BIOLINE NS1 Ag were
Duo rapid test during a
equivalent to Platelia NS1, 471 patients (14.1%) were NS1 Ag positive with the RDT and 14.2% with Platelia. The
dengue virus epidemic
Cohen’s Kappa coefficient was 0.86 *95%CI: 0.83–0.88+, indicating an almost perfect agreement. Moreover, the
sensitivity of SD BIOLINE NS1 Ag relative to the RT-PCR method was 87% *95%CI: 80–93%+ and the specificity was
92% *95% CI: 87–97%+. However, the SD BIOLINE IgM test was found positive in 6.3% of the samples in comparison
to 10.7% with Dx Select IgM. The Cohen’s Kappa coefficient was 0.53 *95%CI: 0.47–0.58+ indicating a moderate
 agreement. This raised concern about the SD BIOLINE IgM for the diagnostic of dengue in endemic areas. When
considering only NS1 Ag results and not IgM, the RDT could be a viable solution to manage dengue outbreaks in
healthcare centers where no laboratory services are available, in the early phase of the disease. © 2017, Springer-
Verlag GmbH Germany.
The recent Zika virus (ZIKV) outbreak demonstrates that cost-effective clinical diagnostics are urgently needed to
detect and distinguish viral infections to improve patient care. Unlike dengue virus (DENV), ZIKV infections during
pregnancy correlate with severe birth defects, including microcephaly and neurological disorders. Because ZIKV and
DENV are related flaviviruses, their homologous proteins and nucleic acids can cause cross-reactions and false-
positive results in molecular, antigenic, and serologic diagnostics. We report the characterization of monoclonal
antibody pairs that have been translated into rapid immunochromatography tests to specifically detect the viral
Rapid antigen tests for nonstructural 1 (NS1) protein antigen and distinguish the four DENV serotypes (DENV1-4) and ZIKV without cross-
dengue virus serotypes reaction. To complement visual test analysis and remove user subjectivity in reading test results, we used image
53.
and zika virus in patient processing and data analysis for data capture and test result quantification. Using a 30-ml serum sample, the
serum sensitivity and specificity values of the DENV1-4 tests and the pan-DENV test, which detects all four dengue
serotypes, ranged from 0.76 to 1.00. Sensitivity/specificity for the ZIKV rapid test was 0.81/0.86, respectively, using a
150-ml serum input. Serum ZIKV NS1 protein concentrations were about 10-fold lower than corresponding DEN NS1
concentrations in infected patients; moreover, ZIKV NS1 protein was not detected in polymerase chain reaction-
positive patient urine samples. Our rapid immunochromatography approach and reagents have immediate
application in differential clinical diagnosis of acute ZIKV and DENV cases, and the platform can be applied toward
developing rapid antigen diagnostics for emerging viruses. © 2017 The Authors, some rights reserved.
OBJECTIVE: To determine the performance of rapid diagnostic tests for dengue and leptospirosis that rely on
detecting antibodies that may not be produced when patients present for medical treatment., METHODS: We
Rapid diagnostic tests for
prospectively enrolled 723 patients with undifferentiated febrile illness presenting to rural hospitals in northern and
dengue and
northeastern Thailand over a 1-year period. We evaluated rapid antibody detection diagnostic tests for dengue and
54. leptospirosis: antibody
leptospirosis on these patients., RESULTS: Sensitivity of the tests was low at the acute visit (7.6-21.5%). Sensitivity at
detection is insensitive at
the convalescent visit ranged from 25.8% to 81.5% and was significantly higher than at the acute visit for all tests
presentation.
([chi]2, P < 0.001)., CONCLUSIONS: Low sensitivity of the rapid tests at presentation suggests that their utility in the
acute phase of dengue and leptospirosis is limited., Copyright (C) 2007 Blackwell Publishing Ltd.
Background Dengue virus (DENV) infection is prevalent across tropical regions and may cause severe disease. Early
diagnosis may improve supportive care. We prospectively assessed the Standard Diagnostics (Korea) BIOLINE Dengue
Rapid Diagnostic Tests
Duo DENV rapid diagnostic test (RDT) to NS1 antigen and anti-DENV IgM (NS1 and IgM) in children in Cambodia, with
for Dengue Virus
the aim of improving the diagnosis of DENV infection. Methodology and principal findings We enrolled children
Infection in Febrile
admitted to hospital with non-localised febrile illnesses during the 5-month DENV transmission season. Clinical and
55. Cambodian Children:
laboratory variables, and DENV RDT results were recorded at admission. Children had blood culture and serological
Diagnostic Accuracy and
and molecular tests for common local pathogens, including reference laboratory DENV NS1 antigen and IgM assays.
Incorporation into
337 children were admitted with non-localised febrile illness over 5 months. 71 (21%) had DENV infection (reference
Diagnostic Algorithms
assay positive). Sensitivity was 58%, and specificity 85% for RDT NS1 and IgM combined. Conditional inference
framework analysis showed the additional value of platelet and white cell counts for diagnosis of DENV infection.
 Variables associated with diagnosis of DENV infection were not associated with critical care admission (70 children,
21%) or mortality (19 children, 6%). Known causes of mortality were melioidosis (4), other sepsis (5), and malignancy
(1). 22 (27%) children with a positive DENV RDT had a treatable other infection. Conclusions The DENV RDT had low
sensitivity for the diagnosis of DENV infection. The high co-prevalence of infections in our cohort indicates the need
for a broad microbiological assessment of non-localised febrile illness in these children. © 2015 Carter et al.
A commercial capture ELISA for specific IgM and IgG antibodies produced during dengue infection (PanBio Dengue
Duo) showed excellent sensitivity (99%, n = 78) using sera collected at hospital discharge compared with established
Rapid serologic diagnosis
ELISA and hemagglutination inhibition (HAI) assays. Furthermore, the ELISA was able to diagnose 79% of the dengue
of dengue virus infection
cases using sera collected at hospital admission. The ELISA also showed high specificity (92%) in paired sera from
using a commercial
56. patients without flavivirus infection (n = 26), although 45% of the patients with Japanese encephalitis (n = 20) showed
capture ELISA that
elevation of IgG but not IgM. The IgG capture ELISA showed good correlation with the HAI assay (r = 0.83, P < 0.0001),
distinguishes primary
and IgG levels could be used to distinguish between primary and secondary infection, with 100% of primary infections
and secondary infections
and 96% of secondary infections being correctly classified. This ELISA should prove useful in the clinical diagnosis of
dengue infections.
Dengue virus (DENV) is the most prevalent arbovirus in the world, found mainly in tropical regions. As clinical
manifestations present frequently as nonspecific febrile illness, laboratory diagnosis is essential to confirm DENV
infections and for epidemiological studies. Recombinant envelope (E) antigens of four serotypes of DENV were used
Recombinant envelope
to develop an immunoglobulin G enzyme-linked immunosorbent assay (IgG-ELISA). To evaluate the IgG-ELISA, a panel
protein-based enzyme
of serum samples that had been tested previously by a plaque reduction neutralization test (PRNT) was investigated
immunoassay for IgG
for the presence of anti-E antibodies against the four DENV serotypes. IgG-ELISA was found to have a sensitivity (91%)
57. antibodies is comparable
and specificity (98%) at a receiver-operating characteristic (ROC) optimized cutoff and demonstrated high
to neutralization tests for
performance as well as good indexes. A concordance of 97% was achieved between both assays, and only 21/704
epidemiological studies
(3%) samples were not concordant. The results of the present study demonstrate a moderate correlation between
of dengue infection
neutralizing antibody titers and IgG-ELISA values. These findings indicate that the recombinant protein-based IgG-
ELISA is a suitable method for routine serodiagnosis, monitoring and seroepidemiological studies of DENV infections.
© 2012 Elsevier B.V.
Dengue fever is a mosquito-borne viral disease prevalent mainly in tropical countries. As the clinical manifestations of
dengue are not very unique, laboratory diagnosis is crucial in identifying cases of dengue infection. Detection of
dengue infection based on the identification of antidengue antibodies has emerged as a practical and reliable means
of diagnosing dengue fever. We recently developed a customized recombinant dengue multiepitope protein (r-DME-
Recombinant G) that can specifically detect the immunoglobulin G (IgG) class of antidengue antibodies in patient sera. Using this
multiepitope protein for strategy, we have now created another dengue multiepitope protein, r-DME-M, with specificity for the IgM class of
58.
early detection of antidengue antibodies. A synthetic gene encoding the r-DME-M protein was expressed as a maltose-binding protein
dengue infections fusion in Escherichia coli. The recombinant protein was purified in a single affinity chromatographic step to obtain
yields of similar to 15 mg purified protein/liter of culture. The purified protein was used to develop an in-house IgM
enzyme-linked immunosorbent assay (ELISA) and tested using a panel of 172 patient sera characterized using the
commercially available Dengue Duo rapid strip test from PanBio, Australia. The IgM ELISA results showed that the r-
DME-M protein not only recognized all IgM(+) samples identified by the PanBio test but also identified samples
 missed by the latter test. We also successfully adapted the r-DME-M protein to a rapid strip test format. This
approach of creating customized antigens coupled to overexpression in E. coli and simple purification offers a
promising alternative option to dengue diagnosis with the potential to circumvent the drawbacks of the whole virus
antigen-based commercial kits.
Dengue is one of the rapidly emerging global threats. In situations of epidemics and routine cases, early diagnosis is
the key to successful management of dengue cases. There are many diagnostic kits available commercially,but their
Seroprevalence and
validity is unknown. The standard test is ELISA. In the present study, the test results of rapid immunochromatographic
evaluation of rapid
test is compared with ELISA. Materials & Method: The study was conducted from December 2012 to August 2014 at
immunochromatographi
Shri B M Patil Medical College & Research centre, Bijapur, Karnataka. Probable dengue cases were diagnosed as per
c test and ELISA for the
the WHO criteria and rapid immunochromatographic test and ELISA were performed on the serum samples for the
detection of NS1 antigen,
59. detection of NS1 antigen, IgM & IgG antibodies. Results were analyzed. Results: A total of 90 cases were enrolled. 36
IgM and IgG antibodies
(40%)cases were found to be positive by rapid immunochromatographic test, whereas 39(43%) cases were found to
for early diagnosis of
be positive by ELISA. The sensitivity of rapid test was 92.31% & specificity was 100% along with positive predictive
dengue infection in a
value of 100% and negative predictive value of 94.44% and was compared with other studies. Conclusion: The study
Tertiary care hospital,
showed that the rapid test can be used as a screening test. Highly suspicious cases should be subjected to
south India
investigations with higher sensitivity & specificity(ELISA), though the results take more time. © 2016, Indian Journal
of Public Health Research and Development. All rights reserved.
A cross-sectional sero-epidemiological study was conducted to determine the prevalence of dengue in Trinidad. Two
commercial rapid test kits, PanBio Dengue Duo IgM and IgG Rapid Strip Test and the Bio-Check Plus Dengue G/M
Cassette Test (Brittney) were used. The immunosorbent assay (ELISA) (FOCUS Technologies, California) was used as
the control. One hundred and twenty five cord blood samples were collected (46 from Mt. Hope Women's Hospital
Seroprevalence of (MH) and 79 from the San Fernando General Hospital (SF)). All blood samples were tested in accordance with the two
dengue in Trinidad using rapid kits and ELISA assay manufacturer's instructions. From 125 cord blood samples, the IgG FOCUS ELISA results
60.
rapid test kits: A cord showed 93.5 and 95% infections at MH and SF, respectively. Whereas the Brittney and PanBio kits showed 10.9 and
blood survey 5.1%, and 26.1 and 50.6% for MH and SF, respectively. Based on the FOCUS ELISA (control) assays, the combined
seroprevalence rate from north and south Trinidad was 94.4%. IgG and IgM sensitivity and specificity levels were
higher in the PanBio than Brittney test kits. The high seroprevalence rates observed in Trinidad are discussed to
stimulate more research to explain this phenomenon and to prevent the Southeast Asian scenario from developing in
the Americas. (c) 2007 Elsevier B.V. All rights reserved.
Background: Serological tests for IgM and IgG are routinely used in clinical laboratories for the rapid diagnosis of
The diagnostic sensitivity dengue and can differentiate between primary and secondary infections. Dengue virus non-structural protein 1 (NS1)
of Dengue Rapid test has been identified as an early marker for acute dengue, and is typically present between days 1-9 post-onset of
assays is significantly illness but following seroconversion it can be difficult to detect in serum. Aims: To evaluate the performance of a
61. enhanced by using a newly developed Panbio® Dengue Early Rapid test for NS1 and determine if it can improve diagnostic sensitivity
combined Antigen and when used in combination with a commercial IgM/IgG rapid test. Methodology: The clinical performance of the
Antibody testing Dengue Early Rapid was evaluated in a retrospective study in Vietnam with 198 acute laboratory-confirmed positive
approach and 100 negative samples. The performance of the Dengue Early Rapid in combination with the IgM/IgG Rapid test
was also evaluated in Malaysia with 263 laboratory-confirmed positive and 30 negative samples. Key Results: In
 Vietnam the sensitivity and specificity of the test was 69.2% (95% CI: 62.8% to 75.6%) and 96% (95% CI: 92.2% to
99.8) respectively. In Malaysia the performance was similar with 68.9% sensitivity (95% CI: 61.8% to 76.1%) and 96.7%
specificity (95% CI: 82.8% to 99.9%) compared to RT-PCR. Importantly, when the Dengue Early Rapid test was used in
combination with the IgM/IgG test the sensitivity increased to 93.0%. When the two tests were compared at each day
post-onset of illness there was clear differentiation between the antigen and antibody markers. Conclusions: This
study highlights that using dengue NS1 antigen detection in combination with anti-glycoprotein E IgM and IgG
serology can significantly increase the sensitivity of acute dengue diagnosis and extends the possible window of
detection to include very early acute samples and enhances the clinical utility of rapid immunochromatographic
testing for dengue. © 2011 Fry et al.
Background: Dengue is an important mosquito-borne viral infection that affects millions of persons worldwide. Early
diagnosis is necessary to effect appropriate management and decrease mortality. Immunochromatographic tests are
advantageous in producing dengue test results within 30 min but these results should be sensitive and specific. In this
The performance of the
study we evaluated the diagnostic performance of the SD BIOLINE Dengue DUO (R) rapid immunochromatographic
SD BIOLINE Dengue DUO
test kit. A panel of 309 dengue and 30 non-dengue single serum samples characterized by using reference enzyme-
(R) rapid
linked immunosorbent assays (ELISAs) was used. These samples were received in the virology laboratory for routine
immunochromatographi
testing during a dengue type 1 outbreak between October to December, 2012. Results: The overall diagnostic
62. c test kit for the
sensitivities of the SD BIOLINE Dengue DUO (R) rapid testfor IgM, IgG and NSI were 49.3 % (95 % CI: 41.3-57.4), 39.1 %
detection of NS1 antigen,
(95 % CI: 33.3-45.2) and 90 % (95 % CI: 82.1-94.7), respectively. The IgM and IgG detection rates were significantly
IgM and IgG antibodies
lower than that of the NSI (p < 0.001). However the combination of the IgM detection with NS1 detection or both NS1
during a dengue type 1
and IgG resulted in a significant (p < 0.001) increase in sensitivity to 97.5 % (95 % CI: 92.9-99.2) and 98.9 % (95 % CI:
epidemic in Jamaica
96.0-99.7), respectively. These higher sensitivities were achieved without any decrease in specificities. Conclusions:
This study revealed that combining two or more parameters of the SD BIOLINE Dengue DUO (R) rapid kit significantly
improved the sensitivity of diagnosis of dengue virus infection and supports its usefulness in the Jamaican setting.
Two cases of false-
positive dengue non-
structural protein 1 (NS1)
Early diagnosis of dengue has been made easier in recent years owing to the advancement in diagnostic technologies.
antigen in patients with
The rapid non-structural protein 1 (NS1) test strip is widely used in many developed and developing regions at risk of
hematological
63. dengue. Despite the relatively high specificity of this test, we recently encountered two cases of false-positive dengue
malignancies and a
NS1 antigen in patients with underlying hematological malignancies.We reviewed the literature for causes of
review of the literature
falsepositive dengue NS1. Copyright © 2015 by The American Society of Tropical Medicine and Hygiene.
on the use of NS1 for the
detection of dengue
infection
Use of a rapid test for Dengue is major public health problem, globally. Timely verification of suspected dengue outbreaks allows for public
diagnosis of dengue health response, leading to the initiation of appropriate clinical care. Because the clinical presentation of dengue is
64. during suspected dengue nonspecific, dengue diagnosis would benefit from a sensitive rapid diagnostic test (RDT). We evaluated the diagnostic
outbreaks in resource- performance of an RDT that detects dengue virus (DENV) nonstructural protein 1 (NS1) and anti-DENV IgM during
limited regions suspected acute febrile illness (AFI) outbreaks in four countries. Real-time reverse transcription-PCR and anti-DENV
 IgM enzyme-linked immunosorbent assay were used to verify RDT results. Anti-DENV IgM RDT sensitivity and
specificity ranged from 55.3 to 91.7% and 85.3 to 98.5%, respectively, and NS1 sensitivity and specificity ranged from
49.7 to 92.9% and 22.2 to 89.0%, respectively. Sensitivity varied by timing of specimen collection and DENV serotype.
Combined test results moderately improved the sensitivity. The use of RDTs identified dengue as the cause of AFI
outbreaks where reference diagnostic testing was limited or unavailable. Copyright © 2016 David et al.
An immunochromatographic test that incorporates recombinant antigens (Dengue Duo Rapid Strip Test; PanBio,
Use of recombinant
Brisbane, Australia) has recently become commercially available. This assay is performed in 15 min and detects both
envelope proteins for
immunoglobulin M (IgM) and IgG in a capture format. The four recombinant proteins used represent the N-terminal
serological diagnosis of
80% of the viral envelope glycoproteins of dengue viruses 1, 2, 3, and 4, respectively. The sensitivity and specificity of
65. dengue virus infection in
the recombinant-antigen-based assay were 90 and 86%, respectively. The similar diagnostic performance of these
an
antigens to that of enzyme-linked immunosorbent assays using whole dengue virus suggests that they mimic whole
immunochromatographi
dengue viruses in primary structure and epitope conformation. These results suggest that recombinant proteins can
c assay
be used in diagnostic assays for dengue to overcome safety issues associated with the use of whole virus.
BACKGROUND The diagnosis of dengue is complex. Until recently, only specialised laboratories were able to confirm
dengue infection. However, this has changed with the newly available immunochromatographic rapid tests. Early
diagnosis is of great interest, and point-of-care rapid tests have been increasingly used in Brazil. Most of those tests
have not undergone validation in the Brazilian population. In this context, we decided to evaluate a rapid test
introduced in the Federal District (FD). OBJECTIVES To estimate the accuracy and reliability of the SD Bioeasy Dengue
Duo rapid test and its components to detect dengue infections in a consecutive sample of symptomatic residents in
the FD, Brazil. METHODS In total, 1353 venous blood samples were collected between 2013 and 2014. Two hundred
Validation and reliability and six positive samples (cases) and 246 negative samples (non cases) were required for sensitivity and specificity
of the rapid diagnostic estimation, respectively; for agreement evaluation, we used 401 samples. The reference standard used was a
test ‘SD bioeasy composite of MAC-ELISA, virus isolation and real-time polymerase chain reaction (RT-qPCR). The evaluation was
66.
dengue duo’ for conducted prospectively under field conditions in the public health units of the FD. FINDINGS The results for the
dengue diagnosis in overall accuracy of the rapid test (NS1/IgM combined) showed 76% sensitivity and 98% specificity. The sensitivity for
Brazil: A phase III study the NS1 component (67%) was better than that for the IgM component (35%). The positive likelihood ratio was 46,
and the negative likelihood ratio was 0.24. The reliability of the test (NS1/IgM combined) demonstrated crude
agreement of 98% (Kappa index 0.94). MAIN CONCLUSIONS The present phase III, large-scale validation study
demonstrates that the rapid test SD Bioeasy Dengue Duo has moderate sensitivity (NS1/IgM combined) and high
specificity. Therefore, the test is useful in confirming the diagnosis of dengue, but not enough to rule out the
diagnosis. Our results also suggest that Dengue virus (DENV) viral load estimated through the RT-qPCR and antibody
level measured through the MAC-ELISA could have had a direct influence on the accuracy of the rapid test. © 2018,
Fundacao Oswaldo Cruz. All rights reserved.
Validation and reliability BACKGROUND The diagnosis of dengue is complex. Until recently, only specialised laboratories were able to confirm
of the rapid diagnostic dengue infection. However, this has changed with the newly available immunochromatographic rapid tests. Early
67. test 'SD Bioeasy Dengue diagnosis is of great interest, and point-of-care rapid tests have been increasingly used in Brazil. Most of those tests
Duo' for dengue have not undergone validation in the Brazilian population. In this context, we decided to evaluate a rapid test
diagnosis in Brazil: a introduced in the Federal District (FD). OBJECTIVES To estimate the accuracy and reliability of the SD Bioeasy Dengue
phase III study Duo rapid test and its components to detect dengue infections in a consecutive sample of symptomatic residents in
the FD, Brazil. METHODS In total, 1353 venous blood samples were collected between 2013 and 2014. Two hundred
and six positive samples (cases) and 246 negative samples (non cases) were required for sensitivity and specificity
estimation, respectively; for agreement evaluation, we used 401 samples. The reference standard used was a
composite of MAC-ELISA, virus isolation and real-time polymerase chain reaction (RT-qPCR). The evaluation was
conducted prospectively under field conditions in the public health units of the FD. FINDINGS The results for the
overall accuracy of the rapid test (NS1/IgM combined) showed 76% sensitivity and 98% specificity. The sensitivity for
the NS1 component (67%) was better than that for the IgM component (35%). The positive likelihood ratio was 46,
and the negative likelihood ratio was 0.24. The reliability of the test (NS1/IgM combined) demonstrated crude
agreement of 98% (Kappa index 0.94). MAIN CONCLUSIONS The present phase III, large-scale validation study
demonstrates that the rapid test SD Bioeasy Dengue Duo has moderate sensitivity (NS1/IgM combined) and high
specificity. Therefore, the test is useful in confirming the diagnosis of dengue, but not enough to rule out the
diagnosis. Our results also suggest that Dengue virus (DENV) viral load estimated through the RT-qPCR and antibody
level measured through the MAC-ELISA could have had a direct influence on the accuracy of the rapid test.