Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul, Korea.
Ann Lab Med. 2017 Nov;37(6):494-498. doi: 10.3343/alm.2017.37.6.494.
Molecular detection of Middle East respiratory syndrome coronavirus (MERS-CoV) using real-time reverse transcription (rRT)-PCR assays is the method of choice for diagnosis of MERS. We evaluated the performance of the PowerChek MERS (upE & ORF1a) real-time PCR Kit (PowerChek MERS assay; Kogene Biotech, Korea) a one-step rRT-PCR assay for the qualitative detection of MERS-CoV.
We evaluated PowerChek MERS assay performance in comparison with nested RT-PCR and sequencing of the RNA-dependent RNA polymerase (RdRp) and N genes. To evaluate diagnostic sensitivity and specificity, 100 clinical specimens (50 positive and 50 negative for MERS-CoV) were simultaneously tested by using the PowerChek MERS and sequencing assays. Assay performance, including limit of detection and precision, was evaluated in vitro by using MERS-CoV RNA transcripts. Analytical specificity was evaluated with a diverse collection of 16 respiratory virus-positive clinical specimens and 14 respiratory bacterial isolates.
The 95% limits of detection of the PowerChek MERS assay for the upE and the open rading frame (ORF)1a were 16.2 copies/μL and 8.2 copies/μL, respectively. No cross-reactivity was observed. The diagnostic sensitivity and specificity of the PowerChek MERS assay were both 100% (95% confidence interval, 91.1-100%).
The PowerChek MERS assay is a straightforward and accurate assay for detecting MERS-CoV RNA. The assay will be a useful tool for the rapid diagnosis of MERS and could prove especially important for MERS outbreak control.
使用实时逆转录（rRT）-PCR 检测中东呼吸综合征冠状病毒（MERS-CoV）的分子检测是诊断 MERS 的首选方法。我们评估了 PowerChek MERS（upE 和 ORF1a）实时 PCR 试剂盒（PowerChek MERS 检测；韩国 Kogene Biotech）的性能，这是一种用于定性检测 MERS-CoV 的一步 rRT-PCR 检测。
我们将 PowerChek MERS 检测与嵌套 RT-PCR 和 RNA 依赖性 RNA 聚合酶（RdRp）和 N 基因测序进行比较，评估其性能。为了评估诊断的敏感性和特异性，我们使用 PowerChek MERS 和测序检测同时对 100 份临床标本（50 份 MERS-CoV 阳性和 50 份阴性）进行检测。通过使用 MERS-CoV RNA 转录物，评估了检测的体外极限检测限和精密度。通过对 16 种呼吸道病毒阳性临床标本和 14 种呼吸道细菌分离株的多样性收集评估了分析特异性。
PowerChek MERS 检测对 upE 和开放阅读框（ORF）1a 的 95%检测限分别为 16.2 拷贝/μL 和 8.2 拷贝/μL。未观察到交叉反应。PowerChek MERS 检测的诊断敏感性和特异性均为 100%（95%置信区间，91.1-100%）。
PowerChek MERS 检测是一种用于检测 MERS-CoV RNA 的简单而准确的检测方法。该检测方法将成为快速诊断 MERS 的有用工具，特别是对 MERS 爆发控制将非常重要。
