![]() ![]() Plates were read with an EL-340 plate reader and KC4 software (BioTek Instruments Winooski, VT, USA). The samples in the dilution plate were then mixed and added to Focus HSV-1 or HSV-2 reaction plates using an eight-channel pipetter. The UWVL HSV-1 internal positive control was added to each HSV-1 plate, and the UWVL HSV-2 positive control was added to each HSV-2 plate. Kit controls, including high positive, low positive, negative and calibrator sera, were also diluted 1 : 101. Each sample was vortexed, and 2.5 μL was diluted 1 : 101 into dilution wells containing 250 μL of sample diluent (Focus Technologies). For clarity, the tests are referred to as Focus-HSV-1 or Focus-HSV-2 in this paper. The name of the tests has since been changed by Focus Technologies to HerpeSelect. Samples were tested in Seattle by HSV-1 ELISA IgG and by HSV-2 ELISA IgG (MRL, now Focus Technologies Cypress, CA, USA). Performance data generated in one area may not be applicable to other populations. In summary, the sensitivity and specificity of the Focus-HSV-1 and Focus-HSV-2 tests varied by site. A sensitivity of 90% and a specificity of 98% were achieved for sites other than Nigeria with the higher cut-off. Raising the positive cut-off index value for HSV-2 from 1.1 to 3.5 yielded a sensitivity of 90% and a specificity of 96%. Focus-HSV-2 sensitivity and specificity in sites other than Ibadan were 97% and 93%, respectively. Concordance of Focus-HSV-2 and WB was 92%, with 97% sensitivity and 89% specificity. Specimens from Songkla, Thailand had 84% concordance with WB results for HSV-1, while three other sites had 100% concordance. Focus-HSV-1 and WB results were concordant (both negative or both positive) for 97% of samples, with 99% sensitivity and 77% specificity. The sensitivity and specificity were determined with samples from ten sites ( n = 967) for Focus-HSV-1 and from seven sites ( n = 675) for Focus-HSV-2. A subset of sera from each site was then selected, based on the HSV-2 results, and tested by Western blot (WB). Sera with Focus-HSV-2-positive results were retested 94% of the 3617 samples retested were positive. To determine the sensitivity and specificity of Focus HerpeSelect ELISAs, sera or plasma samples from women aged 18–55 years were collected in ten cities from eight countries and tested by HerpeSelect HSV-1 ELISA (Focus-HSV-1) and by HerpeSelect HSV-2 ELISA (Focus-HSV-2). ![]()
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