O'Hara Robert William, Brown Benjamin, Hughes Angela, McEwan Ashley, Birtles Andrew, Hawker Adam, Davies Emma, Farooq Hamzah Z, Tilston Peter, Haigh Dominic, Hesketh Louise, Dodgson Andrew, Dodgson Kirsty, Shazaad Ahmad, Guiver Malcolm, Machin Nicholas
Department of Virology, UK Health Security Agency Manchester, Oxford Road, Manchester M13 9WL, UK.
Department of Virology, Manchester Royal Infirmary, Manchester University NHS Foundation Trust, Manchester, UK.
J Clin Virol Plus. 2022 Aug;2(3):100098. doi: 10.1016/j.jcvp.2022.100098. Epub 2022 Jul 16.
Here we describe a retrospective clinical evaluation of the QIAGEN SARS-CoV-2 Prep&Amp UM RT-PCR assay that detects SARS-CoV-2 RNA without the need for a nucleic acid eluate extraction procedure. Using Roche SARS-CoV-2 RT-PCR on the cobas® 8800 platform as a reference standard, a total of 225 confirmed SARS-CoV-2 positive and 320 negative nasopharyngeal swabs in viral transport media, were used to evaluate the assay. Using the RT-PCR cycle threshold as a semi-quantitative marker of viral load, an assessment of over 370,000 SARS-CoV-2 RT-PCR positive results was used in the design of the reference positive specimen cohort. The viral load of all reference positive specimens used in the evaluation was a unique and accurate representation of the range and levels of SARS-CoV-2 positivity observed over a 13-month period of the COVID-19 pandemic. The RT-PCR detects the presence of SARS-CoV-2 RNA, an internal control, and the human RNase P gene to ensure specimen quality. The diagnostic sensitivity of was 92.89% with a specificity of 100%. To assess the analytical sensitivity, a limit of detection was performed using the 1 WHO NIBSC SARS-CoV-2 international standard, recording a 95% LOD of 1.1 × 10 IU/ml. The total invalid rate of specimens was 7.34% due to a lack of detectable RNase P (C >35). The SARS-CoV-2 Prep&Amp UM RT-PCR assay is a new rapid RT-PCR assay, which may be considered to produce acceptable levels of diagnostic sensitivity and specificity whilst potentially halving the laboratory processing time.
在此，我们描述了对QIAGEN SARS-CoV-2 Prep&Amp UM RT-PCR检测法的回顾性临床评估，该检测法无需核酸洗脱提取程序即可检测SARS-CoV-2 RNA。以罗氏在cobas® 8800平台上的SARS-CoV-2 RT-PCR作为参考标准，共使用了225份确诊为SARS-CoV-2阳性和320份阴性的病毒运输培养基中的鼻咽拭子来评估该检测法。以RT-PCR循环阈值作为病毒载量的半定量标志物，在设计参考阳性标本队列时使用了超过370,000份SARS-CoV-2 RT-PCR阳性结果进行评估。评估中使用的所有参考阳性标本的病毒载量是在COVID-19大流行的13个月期间观察到的SARS-CoV-2阳性范围和水平的独特且准确的代表。该RT-PCR检测SARS-CoV-2 RNA、一个内部对照以及人类RNase P基因的存在，以确保标本质量。其诊断敏感性为92.89%，特异性为100%。为评估分析敏感性，使用1份世界卫生组织NIBSC SARS-CoV-2国际标准品进行检测限测定，记录的95%检测限为1.1×10 IU/ml。由于缺乏可检测到的RNase P（C>35），标本的总无效率为7.34%。QIAGEN SARS-CoV-2 Prep&Amp UM RT-PCR检测法是一种新的快速RT-PCR检测法，可能被认为能产生可接受水平的诊断敏感性和特异性，同时可能将实验室处理时间减半。
