Comparison of seven commercial RT-PCR diagnostic kits for COVID-19.
Comparison of seven commercial RT-PCR diagnostic kits for COVID-19.
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DOI：
10.1016/j.jcv.2020.104412
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发表时间：
2020-07-01
期刊：
影响因子：
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通讯作者：
Meijer, Adam
中科院分区：
文献类型：
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作者：
van Kasteren, Puck B;van der Veer, Bas;Meijer, Adam
2019年的最后几个月见证了一种新型冠状病毒在人群中的出现。严重急性呼吸系统综合征冠状病毒2型（SARS-CoV-2）已在地球仪蔓延，给社会造成了重大负担。为减少其传播而采取的措施关键取决于采用最敏感和最具体的方法，即实时逆转录酶聚合酶链反应（RT-PCR），及时和准确地查明病毒感染者。许多商业试剂盒最近已经上市，但其性能尚未得到独立评估。本研究的目的是比较来自七个不同制造商（Altona Diagnostics、BGI、CerTest Biotec、KH Medical、PrimerDesign、R-Biopharm AG和Seegene）的选定RT-PCR试剂盒的基本分析和临床性能。我们使用病毒RNA的系列稀释液来建立PCR效率并估计95%的检测限（L0 D95）。此外，我们运行了一组SARS-CoV-2阳性临床样本（n = 13），以初步评估临床敏感性。最后，我们使用非冠状病毒呼吸道病毒感染阳性的临床样本（n = 6）和一组来自相关人类冠状病毒的RNA来评价检测特异性。所有试验的PCR效率均≥ 96%，估计的LOD 95在6倍范围内变化。使用临床样本，我们观察到试剂盒之间的检出率存在一些差异。重要的是，除SARS-CoV-1 E基因外，所有检测均未显示与其他呼吸道（冠状）病毒的交叉反应性。我们的结论是，本研究中评估的所有RT-PCR试剂盒均可用于经验丰富的分子诊断实验室对患者进行COVID-19的常规诊断。
The final months of 2019 witnessed the emergence of a novel coronavirus in the human population. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has since spread across the globe and is posing a major burden on society. Measures taken to reduce its spread critically depend on timely and accurate identification of virus-infected individuals by the most sensitive and specific method available, i.e. real-time reverse transcriptase PCR (RT-PCR). Many commercial kits have recently become available, but their performance has not yet been independently assessed. The aim of this study was to compare basic analytical and clinical performance of selected RT-PCR kits from seven different manufacturers (Altona Diagnostics, BGI, CerTest Biotec, KH Medical, PrimerDesign, R-Biopharm AG, and Seegene). We used serial dilutions of viral RNA to establish PCR efficiency and estimate the 95 % limit of detection (LOD95). Furthermore, we ran a panel of SARS-CoV-2-positive clinical samples (n = 13) for a preliminary evaluation of clinical sensitivity. Finally, we used clinical samples positive for non-coronavirus respiratory viral infections (n = 6) and a panel of RNA from related human coronaviruses to evaluate assay specificity. PCR efficiency was ≥96 % for all assays and the estimated LOD95 varied within a 6-fold range. Using clinical samples, we observed some variations in detection rate between kits. Importantly, none of the assays showed cross-reactivity with other respiratory (corona)viruses, except as expected for the SARS-CoV-1 E-gene. We conclude that all RT-PCR kits assessed in this study may be used for routine diagnostics of COVID-19 in patients by experienced molecular diagnostic laboratories.