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Analytical sensitivity and efficiency comparisons of SARS-CoV-2 RT-qPCR primer-probe sets.
Nature Microbiology ( IF 20.5 ) Pub Date : 2020-07-10 , DOI: 10.1038/s41564-020-0761-6
Chantal B F Vogels 1 , Anderson F Brito 1 , Anne L Wyllie 1 , Joseph R Fauver 1 , Isabel M Ott 2 , Chaney C Kalinich 1 , Mary E Petrone 1 , Arnau Casanovas-Massana 1 , M Catherine Muenker 1 , Adam J Moore 1 , Jonathan Klein 3 , Peiwen Lu 3 , Alice Lu-Culligan 3 , Xiaodong Jiang 3 , Daniel J Kim 3 , Eriko Kudo 3 , Tianyang Mao 3 , Miyu Moriyama 3 , Ji Eun Oh 3 , Annsea Park 3 , Julio Silva 3 , Eric Song 3 , Takehiro Takahashi 3 , Manabu Taura 3 , Maria Tokuyama 3 , Arvind Venkataraman 3 , Orr-El Weizman 3 , Patrick Wong 3 , Yexin Yang 3 , Nagarjuna R Cheemarla 4 , Elizabeth B White 1 , Sarah Lapidus 1 , Rebecca Earnest 1 , Bertie Geng 5 , Pavithra Vijayakumar 5 , Camila Odio 6 , John Fournier 7 , Santos Bermejo 8 , Shelli Farhadian 7 , Charles S Dela Cruz 8 , Akiko Iwasaki 3, 9 , Albert I Ko 1 , Marie L Landry 4, 7, 10 , Ellen F Foxman 3, 4 , Nathan D Grubaugh 1
Affiliation  

The recent spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) exemplifies the critical need for accurate and rapid diagnostic assays to prompt clinical and public health interventions. Currently, several quantitative reverse transcription–PCR (RT–qPCR) assays are being used by clinical, research and public health laboratories. However, it is currently unclear whether results from different tests are comparable. Our goal was to make independent evaluations of primer–probe sets used in four common SARS-CoV-2 diagnostic assays. From our comparisons of RT–qPCR analytical efficiency and sensitivity, we show that all primer–probe sets can be used to detect SARS-CoV-2 at 500 viral RNA copies per reaction. The exception for this is the RdRp-SARSr (Charité) confirmatory primer–probe set which has low sensitivity, probably due to a mismatch to circulating SARS-CoV-2 in the reverse primer. We did not find evidence for background amplification with pre-COVID-19 samples or recent SARS-CoV-2 evolution decreasing sensitivity. Our recommendation for SARS-CoV-2 diagnostic testing is to select an assay with high sensitivity and that is regionally used, to ease comparability between outcomes.



中文翻译:

SARS-CoV-2 RT-qPCR 引物-探针组的分析灵敏度和效率比较。

最近严重急性呼吸系统综合症冠状病毒 2 (SARS-CoV-2) 的传播表明迫切需要准确和快速的诊断分析,以促进临床和公共卫生干预。目前,临床、研究和公共卫生实验室正在使用几种定量逆转录-PCR (RT-qPCR) 检测方法。然而,目前尚不清楚不同测试的结果是否具有可比性。我们的目标是对四种常见的 SARS-CoV-2 诊断试验中使用的引物-探针组进行独立评估。从我们对 RT-qPCR 分析效率和灵敏度的比较中,我们表明,所有引物-探针组均可用于检测 SARS-CoV-2,每次反应 500 个病毒 RNA 拷贝。例外的是 RdRp-SARSr (Charité) 确认性引物-探针组,它的灵敏度低,可能是由于反向引物与循环的 SARS-CoV-2 不匹配。我们没有发现 COVID-19 之前的样本背景放大或最近 SARS-CoV-2 进化降低灵敏度的证据。我们对 SARS-CoV-2 诊断测试的建议是选择一种灵敏度高且在区域内使用的检测方法,以简化结果之间的可比性。

更新日期:2020-07-10
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