The ongoing coronavirus disease 2019 (COVID-19) pandemic has become a public health emergency. Although many reverse-transcription PCR (RT-PCR) assays have been developed, their performance, especially sensitivity assessment, has been insufficiently tested. In this study, a preliminary comparison of the analytical sensitivity of nine RT-qPCR kits from different manufacturers was first conducted using a certified reference material derived from the genomic RNA of SARS-CoV-2 as the template. Subsequently, three of the nine kits, comprising two highly sensitive kits (DAAN, Huirui) and one less sensitive kit (Geneodx), were selected for further sensitivity and specificity validation. The results revealed variations in the performance between kits of the two groups. For the two highly sensitive kits, the limits of detection at 95 % probability (LOD95%) were 5.6 copies of the N gene and 3.5 copies of the ORF 1ab per reaction (DAAN), and 6.4 (N) and 4.6 (ORF 1ab) copies per reaction (Huirui). These LOD95% values were approximately 3 to 4-fold better than those of the Geneodx Kit. However, none of these three Kits showed cross-reactivity against 6 other types of human coronaviruses or respiratory viruses. Because most of these commercial kits are approved as in vitro diagnostics (testing specimens without direct human contact), it would be beneficial for their manufacturers to improve the diagnostic capability of these kits and thus reduce the clinical risks associated with false-negative results.

持续的 2019 年冠状病毒病 (COVID-19) 大流行已成为突发公共卫生事件。尽管已经开发了许多逆转录 PCR (RT-PCR) 检测方法，但它们的性能，尤其是灵敏度评估，尚未得到充分测试。在这项研究中，首先使用来自 SARS-CoV-2 基因组 RNA 的认证参考材料作为模板，对来自不同制造商的 9 种 RT-qPCR 试剂盒的分析灵敏度进行了初步比较。随后，九个试剂盒中的三个，包括两个高度敏感的试剂盒（DAAN，Huirui）和一个不太敏感的试剂盒（Geneodx）被选择用于进一步的灵敏度和特异性验证。结果揭示了两组试剂盒之间的性能差异。对于两个高度敏感的试剂盒，N基因和每个反应 3.5 个 ORF 1ab 拷贝 (DAAN)，以及每个反应 6.4 ( N ) 和 4.6 (ORF 1ab) 拷贝 (Huirui)。这些 LOD95% 值比 Geneodx 试剂盒的值高大约 3 到 4 倍。然而，这三种试剂盒均未显示出对其他 6 种人类冠状病毒或呼吸道病毒的交叉反应。由于这些商业试剂盒中的大多数都被批准用于体外诊断（在没有直接人类接触的情况下测试标本），因此它们的制造商提高这些试剂盒的诊断能力，从而降低与假阴性结果相关的临床风险将是有益的。