Institute of Hygiene, Cologne Merheim Medical Centre, University Hospital of Witten/Herdecke, Cologne, Germany.
Institute of Pathology, Cologne Merheim Medical Centre, University Hospital of Witten/Herdecke, Cologne, Germany.
J Virol Methods. 2021 Sep;295:114202. doi: 10.1016/j.jviromet.2021.114202. Epub 2021 Jun 2.
In limelight of the ongoing pandemic SARS-CoV-2 testing is critical for the diagnosis of infected patients, contact-tracing and mitigating the transmission. Diagnostic laboratories are expected to provide appropriate testing with maximum accuracy. Real-time reverse transcriptase PCR (RT-PCR) is the diagnostic standard. However, only a handful of studies have reviewed their performance in clinical settings. The aim of this study was to compare the performance of the overall analytical matrix including the extraction kit (BD MAX, Promega, Qiagen), the PCR instrument (Agilent Mx3005 P, BD MAX, Qiagen Rotor-Gene, Roche Cobas z 480) and the RT-PCR assay (Altona Diagnostics, CerTest Biotec, R-Biopharm AG) using predefined samples from proficiency testing organizers. The greatest difference of the cycle threshold values between the matrices was nine cycles. One borderline sample could not be detected by three out of twelve analytical matrices and yielded a false negative result. We therefore conclude that diagnostic laboratories should take the complete analytical matrix in addition to the performance values published by the manufacturer for a respective RT-PCR kit into account. With limited resources laboratories have to validate a wide range of kits to determine appropriate analytical matrices for detecting SARS-CoV-2 reliably. The interpretation of clinical results has to be adapted accordingly.
在当前 SARS-CoV-2 大流行的背景下，检测对于感染患者的诊断、接触者追踪和传播缓解至关重要。诊断实验室应提供具有最大准确性的适当检测。实时逆转录聚合酶链反应（RT-PCR）是诊断标准。然而，只有少数研究回顾了它们在临床环境中的性能。本研究的目的是比较包括提取试剂盒（BD MAX、Promega、Qiagen）、PCR 仪器（Agilent Mx3005 P、BD MAX、Qiagen Rotor-Gene、Roche Cobas z 480）和 RT-PCR 检测（Altona Diagnostics、CerTest Biotec、R-Biopharm AG）在内的整个分析矩阵的性能，使用来自能力验证组织者的预定样本。矩阵之间的循环阈值差异最大为九个周期。有一个边界样本不能被 12 个分析矩阵中的 3 个检测到，导致假阴性结果。因此，我们得出结论，诊断实验室除了制造商为特定 RT-PCR 试剂盒发布的性能值外，还应考虑完整的分析矩阵。由于资源有限，实验室必须验证广泛的试剂盒，以确定可靠检测 SARS-CoV-2 的适当分析矩阵。临床结果的解释必须相应调整。
