Department of Microbiology, Queen Mary Hospital, Hong Kong, China.
Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China.
Biomed Res Int. 2019 Oct 9;2019:5715180. doi: 10.1155/2019/5715180. eCollection 2019.
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® Herpesvirus PCR Kit-has not been well studied. This study evaluated the performance of RealStar® 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® 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® 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.
有几种商业 PCR 试剂盒可用于检测单纯疱疹病毒 (HSV) 和水痘带状疱疹病毒 (VZV)，但一种获得 CE 标志的体外诊断试剂盒——RealStar® Herpesvirus PCR Kit 的检测性能尚未得到充分研究。本研究评估了 RealStar® Herpesvirus PCR Kit 1.0 在 LightCycler® 480 Instrument II 上用于检测和区分人类临床标本中 HSV-1、HSV-2 和 VZV 的性能。我们使用从临床标本中提取的核酸进行连续稀释，评估了 RealStar®和内部多重实时 PCR 检测的分析灵敏度。RealStar®检测的分析灵敏度分别为 HSV-1、HSV-2 和 VZV 的 10、32 和 100 拷贝/反应，略高于内部多重实时 PCR 检测。两种检测的每个病毒靶标循环阈值 (Cp) 值的重复性均令人满意，两种检测的内和间测定变异系数值均低于 5%。使用 153 份临床标本和 15 份能力验证样本评估了 RealStar® Herpesvirus PCR 试剂盒与内部多重实时 PCR 检测相比的诊断性能。与内部检测相比，RealStar®检测的敏感性和特异性均为 100%。RealStar®和内部检测的 Cp 值相关性良好。RealStar® Herpesvirus PCR 是一种敏感、特异和可靠的 HSV-1、HSV-2 和 VZV 检测方法，与实验室开发的检测相比，其验证要求较少。
