Abstract
Several commercial PCR kits are available for detection of herpes simplex virus (HSV) and varicella zoster virus (VZV), but the test performance of one CE-marked in vitro diagnostic kit—RealStar® alpha Herpesvirus PCR Kit—has not been well studied. This study evaluated the performance of RealStar® alpha Herpesvirus PCR Kit 1.0 on the LightCycler® 480 Instrument II for detection and differentiation of HSV-1, HSV-2, and VZV in human clinical specimens. We evaluated the analytical sensitivity of the RealStar® and in-house multiplex real-time PCR assays using serial dilutions of nucleic acids extracted from clinical specimens. The analytical sensitivity of the RealStar® assay was 10, 32, and 100 copies/reaction for HSV-1, HSV-2, and VZV, respectively, which was slightly higher than that of the in-house multiplex real-time PCR assay. Reproducibility of the cycle threshold (Cp) values for each viral target was satisfactory with the intra- and interassay coefficient of variation values below 5% for both assays. One-hundred and fifty-three clinical specimens and 15 proficiency testing samples were used to evaluate the diagnostic performance of RealStar® alpha Herpesvirus PCR Kit against the in-house multiplex real-time PCR assay. The RealStar® assay showed 100% sensitivity and specificity when compared to the in-house assay. Cp values of the RealStar® and in-house assays showed excellent correlation. RealStar® alpha Herpesvirus PCR is a sensitive, specific, and reliable assay for the detection of HSV-1, HSV-2, and VZV, with less extensive verification requirements compared to a laboratory developed assay.
Highlights
Herpes simplex virus- (HSV-) 1, HSV-2, and varicella zoster virus (VZV) are important pathogenic human herpesviruses. e manifestations of HSV-1, HSV-2, and VZV often overlap, making clinical differentiation difficult
In addition to the clinical specimens used for assay validation, 15 samples with various concentrations of HSV/VZV or negative for HSV/VZV from College of American Pathologists (CAP) and Quality Control for Molecular Diagnostics (QCMD) were used for external quality assessment (EQA)
® Performance characteristics of the RealStar alpha Herpesvirus PCR Kit and the in-house multiplex real-time PCR assay for the detection of HSV-1, HSV-2, and VZV DNA were evaluated
Summary
Herpes simplex virus- (HSV-) 1, HSV-2, and varicella zoster virus (VZV) are important pathogenic human herpesviruses. e manifestations of HSV-1, HSV-2, and VZV often overlap, making clinical differentiation difficult. Herpes simplex virus- (HSV-) 1, HSV-2, and varicella zoster virus (VZV) are important pathogenic human herpesviruses. E manifestations of HSV-1, HSV-2, and VZV often overlap, making clinical differentiation difficult. HSV-1 and HSV-2 both cause genital herpes, which can mimic perineal or sacral herpes zoster due to VZV [1]. Precise virological diagnosis is important as antiviral dosages and schedules for HSV-1, HSV-2, and VZV infections differ [2, 3]. Cell culture and direct fluorescence-antibody assay have been used for HSV and VZV detection, but they are less sensitive when compared to molecular tests [4,5,6]. Herpesvirus PCR Kit 1.0 capable of detecting and differentiating HSV-1, HSV-2, and VZV against our in-house developed multiplex PCR assay using archived clinical specimens and proficiency testing samples
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