用于通过逆转录定量聚合酶链反应检测严重急性呼吸综合征冠状病毒2的基于苯酚-氯仿的RNA纯化方法:与自动化系统的比较
Phenol-chloroform-based RNA purification for detection of SARS-CoV-2 by RT-qPCR: Comparison with automated systems.
作者信息
Dimke Henrik, Larsen Sanne L, Skov Marianne N, Larsen Hanne, Hartmeyer Gitte N, Moeller Jesper B
机构信息
Department of Cardiovascular and Renal Research, Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark.
Department of Nephrology, Odense University Hospital, Odense, Denmark.
出版信息
PLoS One. 2021 Feb 24;16(2):e0247524. doi: 10.1371/journal.pone.0247524. eCollection 2021.
The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly reached pandemic levels. Sufficient testing for SARS-CoV-2 has remained essential for tracking and containing the virus. SARS-CoV-2 testing capabilities are still limited in many countries. Here, we explore the use of conventional RNA purification as an alternative to automated systems for detection of SARS-CoV-2 by RT-qPCR. 87 clinical swab specimens were extracted by conventional phenol-chloroform RNA purification and compared to commercial platforms for RNA extraction and the fully integrated Cobas®6800 diagnostic system. Our results show that the conventional RNA extraction is fully comparable to modern automated systems regarding analytical sensitivity and specificity with respect to detection of SARS-CoV-2 as evaluated by RT-qPCR. Moreover, the method is easily scalable and implemented in conventional laboratories as a low cost and suitable alternative to automated systems for the detection of SARS-CoV-2.
严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的爆发迅速达到了大流行程度。对SARS-CoV-2进行充分检测对于追踪和控制该病毒仍然至关重要。在许多国家,SARS-CoV-2的检测能力仍然有限。在此,我们探索使用传统的RNA纯化方法作为自动化系统的替代方法,通过逆转录定量聚合酶链反应(RT-qPCR)检测SARS-CoV-2。通过传统的苯酚-氯仿RNA纯化方法提取了87份临床拭子标本,并与用于RNA提取的商业平台以及完全集成的Cobas®6800诊断系统进行了比较。我们的结果表明,通过RT-qPCR评估,在检测SARS-CoV-2的分析灵敏度和特异性方面,传统RNA提取方法与现代自动化系统完全可比。此外,该方法易于扩展,并且可以在传统实验室中实施,作为检测SARS-CoV-2的自动化系统的低成本且合适的替代方法。