Nasal Swab test :: Nasopharyngeal aspirates:: Both of them for influenza tests.

Obtaining an aspirate is, however, unpleasant, and it requires a suction device, features which make it unfeasible for widespread use in clinical practice. The collection of a nasal swab is easy and painless, and it can be done everywhere without any additional devices. For the nasopharyngeal aspirate, a disposable catheter connected to a mucus extractor was inserted into the left nostril to a depth of 5 to 7 cm and drawn back while applying gentle suction with an electric suction device. Immediately after suctioning of the secretions, a sterile cotton swab was dipped into the aspirate and placed into a vial containing viral transport medium as described above.

A throat swab culture is a laboratory test done to identify germs that may cause infection in the throat. It is most often used to diagnose strep throat. You will be asked to tilt your head back and open your mouth wide. The health care provider rubs a sterile cotton swab along the back of your throat near the tonsils. You need to resist gagging and closing the mouth while the swab touches this area. The health care provider may need to scrape the back of the throat with the swab several times. This helps improve the chances of detecting bacteria. Your throat may be sore at the time the test is taken. You may experience a gagging sensation when the back of your throat is touched with the swab, but the test only lasts a few seconds. This test is safe and well-tolerated. In very few patients, the sensation of gagging may lead to an urge to vomit or cough.

For HIV only ::

1. Particle Agglutination tests: they use different types of particles to produce clumping or settling patterns of the particles when a specimen is positive. An autologous red cell agglutination method detects HIV antibodies with a hybrid antigen-antibody reagent which agglutinates the patient's red blood cells. A latex particle agglutination detects HIV antibodies by the agglutination of minute latex particles when mixed with the patients blood. A newer method uses fluid capillary action to enhance and quicken particle agglutination. Particle adherence detects HIV antibodies when the settling pattern of small gelatin particles is altered. (Examples: Capillus, Serodia )

Negative Point ::: a).Serum or plasma is used in the test. ( Serodia ) Blood mostly not preferred. I cite Human serum is the specimen of choice for this kit (4), however EDTA, Sodium Citrate or Heparin plasma may be used when serum cannot be obtained b). Some reagents contain small amounts of sodium azide as preservative. Sodium azide may react with lead or copper plumbing, which may result in the formation of highly explosive metal azides. If these reagents are to be disposed of in a laboratory sink, flush with generous amounts of water to avoid azide build-up. c). Separate preparation of reagents for testing. d). Particle agglutination assays typically require 10 to 60 minutes or more and must be used with serum or plasma. When a patient specimen containing HIV antibodies is mixed with minute HIV antigen-coated latex particles, cross linking occurs and results in agglutination. Some devices enhance the visual agglutination reaction by using small, channeled, clear plastic cassettes. Flow of the specimen-particle mixture through narrowed areas in the channels promotes agglutination. Detection of weak agglutination can be difficult, and readers have been developed for some tests to reduce the inaccuracy introduced by subjective interpretation. The reagents often require refrigeration, and costs range from Rs.120 to Rs.240 per test. ( Reference : http://www.fdi.com/documents/products/inserts/PROOF%20POLLOCK%20Serodia_TP_PA_Sheet%2 0020912.pdf )

2. Immunofiltration Process or Flow through cassettes, or membrane immunoconcentration devices: they capture and detect HIV antibody in a specimen flowing through a porous membrane. A visible dot or line forms on the membrane when HIV antibodies are present. (Examples: Multi-spot, Genie II). Solid phase tests include the dipstick "comb" assay. This assay uses a solid plastic matrix to which an HIV antigen is fixed. When HIV antibodies are present, a spot or dot will be visible when processed with a signal reagent.

Negative points :: Membrane immunoconcentration devices employ solid-phase capture technology, which involves the immobilization of HIV antigens on a porous membrane. The specimen flows through the membrane and is absorbed into an absorbent pad. A dot or a line visibly forms on the membrane when developed with a signal reagent (usually a colloidal gold or selenium conjugate). Some tests allow the differentiation of HIV-1 from HIV-2 by applying antigens from these viruses to different sites on the membrane: The flow-through tests require several steps for the addition of specimen, wash buffers, and signal reagent, and they can usually be performed in 5 to 15 minutes. Most are used with serum or plasma, though some are equipped with a filter to allow the use of whole-blood specimens. The devices or reagents typically require refrigeration. Costs range from Rs.240 to rs.480 per test. However, the assay format has become more rare since the introduction of large numbers of immunochromatographic devices that are considerably simpler to use.

There are also two other formats which are rarely used. Autologous red-cell agglutination tests require 5 minutes or less and detect HIV antibodies with a hybrid antigen-antibody reagent, which, when added to the red cells of the patient, agglutinates the patient's own red cells. Immunodot comb assays use a solid plastic matrix with "teeth" attached to one another, to which HIV antigen is fixed to capture HIV antibodies. Patient specimens are placed in wells spaced to accommodate each tooth of the comb device, which allows batch processing. The tests, which require less than 30 minutes to perform, are then developed with a signal reagent. Results for each specimen are visualized as a spot or a dot on the corresponding tooth.

5. Optical immunoassays All assays in the group of manual optical immunoassays are offered by Thermo Electron Corporation under the branch name OIA Optical ImmunoAssay. The assays are based on a patented thin film technology that utilizes the reflective properties of light from optically coated surfaces. Because a change in reflected color can be directly detected via the antibodyantigen reaction, the OIA devices can be classified as optical immunosensors, i.e., devices that can directly convert a physical signal into an optical one . However, to increase the assay sensitivities, an enzyme label is employed to produce a precipitating end product. The OIA devices are currently the only manual immunosensors on the market. Despite the complex manual assay procedures , the 530 min assays are only qualitative or semiquantitative in nature. However, even picomolar assay sensitivities have been reported directly in whole blood .

Negative points :: Complexity and high cost.

6. Immunosensors : immunosensors are miniaturized analytical devices that incorporate a recognition element (transducer) capable of converting a surface-bound antibodyantigen interaction into a quantifiable signal . Among the several different types of transducers available, especially those based on electrochemical or optical detection have been the most successful . In principle, both types can either be run as label-free (direct) or label-using (indirect) immunosensors and, similar to other immunoassay methods, they are also divided into heterogeneous and homogeneous assay formats according to the requirement for washing. Negative Points :: The two major challenges for especially label-free and homogeneous optical immunosensors seem to be the non-specific binding and the interference in detection when complex biological matrices such as whole blood are used .

7.Solid Phase Dipsticks :: The tests are comprised of solid, nonporous supports onto which analyte capture molecules are immobilized. Some assays simply require the user to dip the test into a specimen and then wait for a color change indicating the test result. To perform most assays of this type, the dipsticks are incubated with patient specimens followed by an initial wash step, addition of signal reagent, and a final wash step An advantage of solid-phase tests is that they allow individual patients to be tested for one or multiple parameters with a single assay.

Negative points :: Most tests can be completed in one hour or less.Complicated Steps involved.Cannot be automated.

8. Nasal Swab test :: Nasopharyngeal aspirates:: Both of them for influenza tests. Obtaining an aspirate is, however, unpleasant, and it requires a suction device, features which make it unfeasible for widespread use in clinical practice. The collection of a nasal swab is easy and painless, and it can be done everywhere without any additional devices. For the nasopharyngeal aspirate, a disposable catheter connected to a mucus extractor was inserted into the left nostril to a depth of 5 to 7 cm and drawn back while applying gentle suction with an electric suction device. Immediately after suctioning of the secretions, a sterile cotton swab was dipped into the aspirate and placed into a vial containing viral transport medium as described above.

9.A throat swab culture is a laboratory test done to identify germs that may cause infection in the throat. It is most often used to diagnose strep throat. You will be asked to tilt your head back and open your mouth wide. The health care provider rubs a sterile cotton swab along the back of your throat near the tonsils. You need to resist gagging and closing the mouth while the swab touches this area. The health care provider may need to scrape the back of the throat with the swab several times. This helps improve the chances of detecting bacteria. Your throat may be sore at the time the test is taken. You may experience a gagging sensation when the back of your throat is touched with the swab, but the test only lasts a few seconds. This test is safe and well-tolerated. In very few patients, the sensation of gagging may lead to an urge to vomit or cough.

Positive points of ImmunoChromatographic tests : Immunochromatographic strips, the most recent development, potentially require only one step and incorporate both antigen and signal reagent into a nitrocellulose strip. The specimen is applied to an absorbent pad from which it is wicked, combines with signal reagent, and migrates through the strip. A positive reaction results in a visual line on the membrane where HIV antigen has been applied. A few of the strip tests also deploy different antigens at different locations to allow differentiation of HIV-1 group M, HIV-1 group O, and HIV-2 antibodies. A procedural control line that detects immunoglobulin G is usually applied to the strip beyond the HIV-antigen line. A visual line at the test and control sites indicates a positive test result, a line only at the control location indicates a negative test result, and the absence of a line at the control site means the test is invalid. Most lateral-flow tests require no additional equipment or refrigeration, and test results can be obtained in less than 15 minutes. Many can be used with whole blood, serum, or plasma, and some can be used with finger-stick specimens, saliva or oral fluids. In some lateral-flow devices, the test strip is encased in a plastic cartridge. Cost of these tests is usually less than Rs.120.

Important paper for support : http://www.banglajol.info/index.php/JAFMC/article/viewFile/8617/6434

References :: http://www.unicef.org/supply/files/HIV_DIAGNOSIS_A_Guide_for_Selecting_RDT_Jan08.pdf http://www.medadvocates.org/diagnostics/cdc/rapidtest.html http://www.testingmillions.org/files/TestingModule2.pdf http://www.bvgh.org/Biopharmaceutical-Solutions/Global-HealthPrimer/Targets/cid/ViewDetails/ItemID/16.aspx http://www.sciencedirect.com/science/article/pii/S0009912005000895

Lateral flow tests are the simplest type of RDT, requiring only very minimal familiarity with the test and no equipment to perform, since all of the reactants and detectors are included in the test strip. In a lateral flow test, the sample is placed into a sample well and migrates across the zone where the antigen or antibody is immobilized. The results are read after a certain amount of time has passed.

Another type of RDT, a flow-through test, obtains results even faster than lateral flow tests, but requires an added wash and buffer step, which can limit its portability and stability.

An agglutination test works very simply by observation of the binding of carrier particles and target analytes into visible clumps, seen either through a microscope or with the naked eye. However, if the binding of the particles is weak, the results of the test can be inconclusive.

Dipstick format RDTs (with binding sites to test for multiple antigens) work by placing the dipstick in a sample. The dipstick is then washed and incubated to prevent non-specific analyte binding. These additional steps can limit their usability in low-resource point of care settings.

Microfluidics, or labs on a chip are an emerging area of rapid diagnostic development. Using electrochemical sensors, these tests would include all detectors and reactants in a single portable cassette.

References for PPT :

*** Case Study ::http://www.bvgh.org/LinkClick.aspx?fileticket=-a1C6u2LE4w%3d&tabid=91 http://www.sciencedirect.com/science/article/pii/S0009912005000895

Might be important :::

Barriers to use of RDT

The main barriers and constraints to the use of rapid diagnostic tests can be put into three main categories:

Acceptability: Rapid tests need to be acceptable to policymakers, clinicians, and patients. Tests need to have sufficient sensitivity and specificity and they need to have an adequate predictive value. Ease-of-use is critical for point-of-care use by clinicians. Culturally appropriate specimens and credible results are important if rapid tests are to be accepted by patients.

Test

Strengths

Weaknesses

Examples

Affordability: Many rapid diagnostic tests are more expensive than the tests or syndromic algorithms they are intended to replace. Decreasing per-test costs, carefully designing diagnostic algorithms, and educating end users about the cost-savings of more efficient use of therapeutic drugs are important means of maximizing rapid test affordability.

Availability: Rapid diagnostic tests are not always available in developing countries. Most tests have limited shelf lives, and many countries have weak public- and private-sector procurement and distribution systems. The consistency and quality of imported tests can also be issues. To address these constraints, local government regulations, quality assurance, shelf life testing, and distribution systems all need to be assessed and improved.

Rapid (5-15 min) Lateral flow tests (immunochromatographic strip tests) Can be adapted for multiple sample types Easiest to use Results are qualitative Less sensitive than an ELISA Malaria RDTs, home pregnancy tests

Low sensitivity and ambiguous Single-step Agglutination assays Low cost per test Rapid results results in weak reactions Some tests require training and/or a microscope to read results Some cross-reactions can cause sensitivity problems

HIV latex agglutination assays, Leishmaniasis DAT

Requires more training to Flow-through Very rapid (3-5 min) perform than lateral flow Less sensitive at antigen detection compared to lateral flow E. coli detection

Solid-phase (dipstick assays)

One strip can test for multiple parameters

Requires several intermediate steps, some training

HIV comb test

Microfluidic chips, immunosensors, labs on a chip

Rapid Requires no manipulation

Potentially prohibitively expensive

Largely hypothetical at this point in time