EVALUATION OF A NONISOTOPIC POLYMERASE CHAIN-REACTION ASSAY FOR DETECTION IN CLINICAL SPECIMENS OF HERPES-SIMPLEX VIRUS TYPE-2 DNA

Background: PCR assays for detection of herpes simplex virus DNA seque nces in clinical specimens are more sensitive than cell culture. Objec tive: A non-isotopic PCR assay (glycoprotein B HSV-2 assay, Roche Mole cular Systems) for detection of HSV-2 DNA sequences was evaluated on 2 34 clinical specimens. Study design: A total of 125 patients (73 women , 40 men, 12 unknown) provided 162 samples contained in viral transpor t medium. Samples were first inoculated in cell culture, centrifuged a t 12 000 x g, lyzed and kept frozen. A total of 77 women provided 42 c ervicovaginal lavages and 30 vaginal tampons that were lyzed, tested w ith two isotopic HSV-2 PCR assays and kept frozen. All these samples w ere subsequently thawed and amplified with the glycoprotein B HSV-2 as say using generic primers for HSV glycoprotein B gene. Amplicons were captured on microplates with a HSV-2-specific probe and were detected with avidin-peroxidase and substrate. Results: Of the 162 samples subm itted to viral culture, HSV-2 was isolated from 73 while 89 did not co ntain HSV-2. All the 73 specimens with culture-proven HSV-2 infections tested positive with glycoprotein B HSV-2 assay (sensitivity of 100%) . Herpesviruses other than HSV-2 were isolated from 34 samples that we re negative with glycoprotein B HSV-2 assay. Two culture-negative samp les tested positive in the glycoprotein B HSV-2 assay (specificity of 98.7%). The latter samples could not be retested in confirmatory isoto pic HSV-2 PCR tests. HSV-2 DNA sequences could also be detected direct ly in cervical lavages or vaginal tampons from 13 women with the glyco protein B HSV-2 assay. Conclusion: Detection and typing of HSV-2 in cl inical samples, including those collected in viral transport medium, c an be accomplished with PCR assays using the AMPLICOR format. (C) 1997 Elsevier Science B.V.