Evaluation of HSV-1 & HSV-2 IgG using the DiaSorin Liaison XL vs Focus Diagnostics HerpeSelect

Bruce
1 Wong ,

Mark
1

1 Vidler ,

Louise

1 Simonetta ,

Noel

1 Fisher ,

Archie

1 Darbar

Pacific Medicine Laboratory Services, Serology, RNSH, NSW (Pathology North)

Introduction
Herpes Simplex Virus 1 and 2 are both members of the Herpesviridae family of viruses. Often called Human Herpes Virus 1 and 2 due to their ability to manifest in human subjects. In Australia the seroprevalence of HSV-1 was 76.5% and HSV-2 was 12 % in 2000. Globally the prevalence of HSV is around 65-90%. The current gold standard diagnostic test for confirming HSV infections is the Western Blot Immunoassay which detects the presence of antibodies produced in response to infection with the virus. Unfortunately this is a labour intensive assay requiring expertise. Most laboratories use an Enzyme-linked immunosorbent assay (ELISA) to determine the presence of type-specific glycoproteins gG1 and gG2 (for HSV-1 and HSV-2 respectively). We have had exposure to 2 assays in our Laboratory, one being the HerpeSelect kit produced by Focus Diagnostics and the other is DiaSorin HSV-1 & HSV-2 kits for the Liason XL. The use of the Focus and DiaSorin assays have allowed laboratories to detect the presence or absence of human IgG class antibodies to either HSV-1 or HSV-2 in human sera and assist diagnosis of past HSV infection in sexually active adults or expectant mothers.

Results
Focus Diagnostics HerpeSelect 1 IgG vs DiaSorin Liaison XL HSV-1 IgG Performance DiaSorin Liaison XL HSV-2 Performance Summary 1.Out of a total of 91 stored serum, 18 samples yielded a positive result and concurred with both the Focus Diagnostics HerpeSelect 2 IgG and the DiaSorin Liaison XL HSV-2 IgG. 2.67 Samples yielded a Negative result and concurred with both the Focus Diagnostics HerpeSelect 2 IgG and the DiaSorin Liaison XL HSV2 IgG. 3.3 Samples on the Focus Diagnostics HerpeSelect 2 IgG assay that were reported Positive, yielded a Negative result on the DiaSorin Liaison XL HSV-2 IgG. 4.3 Sample reported Equivocal on the Focus Diagnostics HerpeSelect 2 IgG yielded a Negative result on the DiaSorin Liaison XL HSV-2 IgG. 5.Agreement with our current method being the Focus Diagnostics HerpeSelect 2 IgG vs the DiaSorin Liaison XL HSV-2 IgG excluding equivocal results was 97%.

Focus Diagnostics HSV-1 IgG

Liaison Positive Equivocal Negative XL HSV-1 IgG Positive 51 3

Total

54

Equivocal

Negative

32

35

Table 1: Results obtained for patients with or without exposure to HSV-1. DiaSorin Liaison XL HSV-1 Performance Summary 1.Out of a total of 91 stored serum, 51 samples yielded a Positive result and concurred with both the Focus Diagnostics HerpeSelect 1 IgG and the DiaSorin Liaison XL HSV-1 IgG. 2.32 Samples on the Focus Diagnostics HerpeSelect 1 IgG assay yielded a Negative result and concurred with both the Focus Diagnostics HerpeSelect 1 IgG and the DiaSorin Liaison XL HSV-1 IgG. 3.2 Samples on the Focus Diagnostics HerpeSelect 1 IgG assay that were reported Positive, yielded an Equivocal result on the DiaSorin Liaison XL HSV-1 IgG. 4.2 Samples on the Focus Diagnostics HerpeSelect 1 IgG assay that were reported Positive, yielded a Negative result on the DiaSorin Liaison XL HSV-1 IgG. 5.3 Sample reported Negative on the Focus Diagnostics HerpeSelect 1 IgG assay yielded a Positive result on the DiaSorin Liaison XL HSV-1 IgG. 6.1 Sample reported Equivocal on the Focus Diagnostics HerpeSelect 1 IgG yielded a Negative result on the DiaSorin Liaison XL HSV-1 IgG. 7.Agreement with our current method being the Focus Diagnostics HerpeSelect 1 IgG vs the DiaSorin Liaison XL HSV-1 IgG excluding equivocal results was 94%.

Conclusion
1. Our results are in broad agreement when we tested and evaluated the Focus Diagnostics HerpeSelect vs DiaSorin Liaison XL HSV assays. 2. Discordant samples should be retested and follow up samples tested should they be available. 3. The gold standard diagnostic test being the HSV Western Blot should be tested on the discordant samples to determine which assay is more sensitive and specific. 4. Further information should be sought about the patients clinical history including any current viral infection which may assist or aid in the diagnosis of HSV infection. 5. For Laboratories considering taking up the DiaSorin Liaison XL Platform for Herpes Simplex Virus testing, the DiaSorin Liaison XL Platform is a suitable alternative to the Focus Diagnostics HerpeSelect assay. 6. Currently, our Laboratory is undertaking further follow up and confirmation using the HSV Western Blot for the discordant samples.

Aim
This evaluation was to determine the performance of the DiaSorin Liason XL in comparison to the Focus Diagnostics HerpeSelect assay, which is currently our method of testing at the Royal North Shore Hospital in St Leonards, New South Wales, Australia.

Method
91 stored adult patients serum were included in the study. All specimens have been previously tested for HSV-1 and HSV-2 using 2 separate Focus Diagnostics HerpeSelect kits being the HerpeSelect 1 and HerpeSelect 2 ELISA IgG assays. All specimens were retested using the DiaSorin Liason XL indirect chemiluminescence immunoassay (CMIA) analyser. The IgG levels were automatically retrieved and analysed by published methods and cut-offs specified by DiaSorin. Calculations are expressed as a qualitative index value, allowing the results and serological characterisation into one of three diagnostic categories: 1. Positive Result: HSV-1 and 2 IgG Index with a value of 1.1 2. Equivocal Results: HSV-1 and 2 IgG Index with a value of 0.9 - 1.1 3. Negative Results: HSV-1 IgG and HSV-2 IgG Index with a value <0.9 Equivocal results require re-testing for a definitive diagnosis and another sample should be submitted to the Laboratory. A second sample should be collected no less than 1 week later. All assays were performed in accordance to the manufacturers instructions. The results of this assay are not diagnostic themselves, and should be correlated with clinical findings and other diagnostic procedures as well as medical judgment.

Focus Diagnostics HerpeSelect 2 IgG vs DiaSorin Liaison XL HSV-2 IgG Performance

Focus Diagnostics HSV-2 IgG

References
A L Cunningham, R Taylor, J Taylor, C Marks, J Shaw, A Mindel , (2006) Prevalence of infection with herpes simplex virus types 1 and 2 in Australia: a nationwide population based survey Sex Transm Infect 82:164-168

Liaison Positive Equivocal Negative XL HSV-2 IgG Positive 18

Total

18

Equivocal

Acknowledgements
PaLMS, a division of Pathology North would like to thank the assistance of DiaSorin for their assistance in the training of Serology Laboratory staff as well as their assistance throughout the evaluation.

Negative

67

73

Table 2: Results obtained for patients with or without exposure to HSV-2.