Department of Laboratory Medicine, Korea University College of Medicine, Seoul, Korea.
J Clin Lab Anal. 2022 Feb;36(2):e24242. doi: 10.1002/jcla.24242. Epub 2022 Jan 12.
Currently, SARS-CoV-2 RNA detection using real-time reverse-transcription PCR (rRT-PCR) is the standard diagnostic test for COVID-19 infection. Various rRT-PCR assays are currently used worldwide, targeting different genes of the SARS-CoV-2. Here, we compared the analytical sensitivity and clinical performance (sensitivity and specificity) of Allplex SARS-CoV-2/FluA/FluB/RSV assay (Seegene), Standard M nCoV real-time detection kit (SD Biosensor), and U-TOP COVID-19 detection kit (Seasun Biomaterials) for SARS-CoV-2 detection.
Two hundred and forty-nine nasopharyngeal swab samples were evaluated to compare the clinical performance of the rRT-PCR assays. For the analytical performance evaluation, two RNA controls with known viral loads-SARS-CoV-2 RNA control and SARS-COV-2 B.1.351 RNA control-were used to investigate the potential impact of SARS-CoV-2 variants, particularly the B.1.351 lineage.
Limits of detection ranged from 650 to 1300 copies/ml for rRT-PCR assays, and the mean differences in cycle threshold (C ) values of the two RNA controls were within 1.0 for each target in the rRT-PCR assays (0.05-0.73), without any prominent C value shift or dropouts in the SARS-COV-2 B.1.351 RNA control. Using the consensus criterion as the reference standard, 89 samples were positive, whereas 160 were negative. The overall clinical performance of rRT-PCR assays was comparable (sensitivity 98.88%-100%; specificity 99.38%-100%), whereas the sensitivities of each target gene were more variable.
The three rRT-PCR assays showed comparable analytical sensitivity and clinical performance. The analytical and clinical sensitivities of each target gene were influenced more by the primer and probe design than the target gene itself.
目前，使用实时逆转录聚合酶链反应（rRT-PCR）检测 SARS-CoV-2 RNA 是 COVID-19 感染的标准诊断测试。目前，世界各地使用了各种 rRT-PCR 检测方法，针对 SARS-CoV-2 的不同基因。在这里，我们比较了 Allplex SARS-CoV-2/FluA/FluB/RSV 检测试剂盒（Seegene）、标准 M nCoV 实时检测试剂盒（SD Biosensor）和 U-TOP COVID-19 检测试剂盒（Seasun Biomaterials）在 SARS-CoV-2 检测中的分析灵敏度和临床性能（敏感性和特异性）。
评估了 249 份鼻咽拭子样本，以比较 rRT-PCR 检测方法的临床性能。对于分析性能评估，使用两个具有已知病毒载量的 RNA 对照 - SARS-CoV-2 RNA 对照和 SARS-COV-2 B.1.351 RNA 对照 - 来研究 SARS-CoV-2 变体，特别是 B.1.351 谱系的潜在影响。
rRT-PCR 检测的检测限范围为 650 至 1300 拷贝/ml，两种 RNA 对照的 C 值差异平均值在 rRT-PCR 检测的每个靶标中均在 1.0 以内（0.05-0.73），在 SARS-COV-2 B.1.351 RNA 对照中没有明显的 C 值偏移或缺失。使用共识标准作为参考标准，89 个样本为阳性，160 个样本为阴性。rRT-PCR 检测的总体临床性能相当（敏感性 98.88%-100%；特异性 99.38%-100%），而每个靶基因的敏感性则更为多变。
三种 rRT-PCR 检测方法显示出相当的分析灵敏度和临床性能。每个靶基因的分析和临床敏感性受引物和探针设计的影响大于靶基因本身。
