Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, South Korea.
Department of Laboratory Medicine, Kangbuk Samsung Hospital, Sungkyunkwan University School of Medicine, Seoul, South Korea.
Microbiol Spectr. 2024 Apr 2;12(4):e0424923. doi: 10.1128/spectrum.04249-23. Epub 2024 Mar 7.
The aim of this study was to compare the performance of the newly developed SMG HHV-6 Q Real-Time PCR Kit (SMG assay) with the RealStar HHV-6 PCR Kit (RealStar assay). The analytical sensitivity and specificity, linearity, and precision of the SMG assay were evaluated. The clinical performance of the SMG assay was assessed and compared with that of the RealStar assay using 207 clinical specimens (HHV-6A positive, = 51; HHV-6B positive, = 64; HHV-6A/B negative, = 92). The limit of detection of the SMG assay was 2.92 log copies/mL for HHV-6A DNA and 2.88 log copies/mL for HHV-6B DNA. The linear range was determined to be 3.40-9.00 log copies/mL for both viruses. Intra- and inter-assay variability were below 5% at concentrations ranging from 4 to 9 log copies/mL. No cross-reactivity was observed with the 25 microorganisms included in the specificity panel. The clinical sensitivity and specificity of the SMG and RealStar assays compared to in-house polymerase chain reaction and sequencing were as follows: SMG assay, 98.0% and 100% for HHV-6A DNA, respectively, and 96.9% and 100% for HHV-6B DNA, respectively; RealStar assay, 98.0% and 100% for HHV-6A DNA, respectively, and 90.6% and 100% for HHV-6B DNA, respectively. The correlation coefficients between viral loads measured by the two assays were 0.948 and 0.975, with mean differences of 0.62 and 0.32 log copies/mL for HHV-6A and HHV-6B DNA, respectively. These results demonstrate that the SMG assay is a sensitive and reliable tool for the quantitative detection and differentiation of HHV-6A and HHV-6B DNA.IMPORTANCEQuantitative real-time PCR (qPCR) that can distinguish between HHV-6A and HHV-6B DNA is recommended for diagnosis of active infection. The SMG HHV-6 Q Real-Time PCR Kit (SMG assay) is a newly developed qPCR assay that can differentiate between HHV-6A and HHV-6B DNA; however, little is known about its performance. In this study, we assessed the performance of the SMG assay and compared it with that of a commercially available qPCR assay, the RealStar HHV-6 PCR Kit (RealStar assay). The SMG assay demonstrated excellent analytical sensitivity and specificity, precision, and linearity. Furthermore, the viral loads measured by the SMG assay were highly correlated with those measured by the RealStar assay. Our results suggest that the SMG assay is a useful diagnostic tool for quantitative detection and differentiation of HHV-6A and HHV-6B DNA.
本研究旨在比较新开发的 SMG HHV-6 Q 实时 PCR 试剂盒（SMG 检测法）与 RealStar HHV-6 PCR 试剂盒（RealStar 检测法）的性能。评估了 SMG 检测法的分析灵敏度和特异性、线性度和精密度。使用 207 份临床标本（HHV-6A 阳性， = 51；HHV-6B 阳性， = 64；HHV-6A/B 阴性， = 92）评估了 SMG 检测法的临床性能，并与 RealStar 检测法进行了比较。SMG 检测法对 HHV-6A DNA 的检测限为 2.92 log 拷贝/mL，对 HHV-6B DNA 的检测限为 2.88 log 拷贝/mL。两种病毒的线性范围均为 3.40-9.00 log 拷贝/mL。在 4 至 9 log 拷贝/mL 的浓度范围内，室内和室内变异系数均低于 5%。在特异性面板中包含的 25 种微生物中未观察到交叉反应。SMG 和 RealStar 检测法与内部聚合酶链反应和测序相比的临床灵敏度和特异性如下：SMG 检测法，HHV-6A DNA 分别为 98.0%和 100%，HHV-6B DNA 分别为 96.9%和 100%；RealStar 检测法，HHV-6A DNA 分别为 98.0%和 100%，HHV-6B DNA 分别为 90.6%和 100%。两种检测法测量的病毒载量之间的相关系数分别为 0.948 和 0.975，HHV-6A 和 HHV-6B DNA 的平均差异分别为 0.62 和 0.32 log 拷贝/mL。这些结果表明，SMG 检测法是一种敏感且可靠的工具，可用于定量检测和区分 HHV-6A 和 HHV-6B DNA。
重要性：
推荐使用能够区分 HHV-6A 和 HHV-6B DNA 的定量实时 PCR（qPCR）来诊断活动性感染。SMG HHV-6 Q 实时 PCR 试剂盒（SMG 检测法）是一种新开发的 qPCR 检测法，可区分 HHV-6A 和 HHV-6B DNA；然而，关于其性能的信息却很少。在这项研究中，我们评估了 SMG 检测法的性能，并将其与市售的 qPCR 检测法 RealStar HHV-6 PCR 试剂盒（RealStar 检测法）进行了比较。SMG 检测法表现出出色的分析灵敏度和特异性、精密度和线性度。此外，SMG 检测法测量的病毒载量与 RealStar 检测法测量的病毒载量高度相关。我们的结果表明，SMG 检测法是一种用于定量检测和区分 HHV-6A 和 HHV-6B DNA 的有用诊断工具。
