Laboratory of Pharmacogenetics, Drug Research and Development Center (NPDM), Federal University of Ceará, Fortaleza, Brazil.
Department of Biochemistry and Molecular Biology (DBBM), Federal University of Ceará, Fortaleza, Brazil.
Expert Rev Mol Diagn. 2022 Feb;22(2):157-167. doi: 10.1080/14737159.2022.2037425. Epub 2022 Feb 21.
The Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) disease has had a catastrophic impact on the world resulting in several deaths. Since World Health Organization declared the pandemic status of the disease, several molecular diagnostic kits have been developed to help the tracking of viruses spread.
This review aims to describe and evaluate the currently reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) diagnosis kit. Several processes used in COVID-19 diagnostic procedures are detailed in further depth to demonstrate optimal practices. Therefore, we debate the main factors that influence the viral detection of SARS-COV-2 and how they can affect the diagnosis of patients.
Here is highlighted and discussed several factors that can interfere in the RT-PCR analysis, such as the viral load of the sample, collection site, collection methodology, sample storage, transport, primer, and probe mismatch/dimerization in different brand kits. This is a pioneer study to discuss the factor that could lead to the wrong interpretation of RT-qPCR diagnosis of SARS-CoV-2. This study aimed to help the readers to understand what very likely is behind a bad result of SARS-CoV-2 detection by RT-PCR and what could be done to reach a reliable diagnosis.
严重急性呼吸系统综合征冠状病毒 2（SARS-CoV-2）疾病对世界造成了灾难性的影响，导致了许多人死亡。自从世界卫生组织宣布该疾病为大流行状态以来，已经开发了几种分子诊断试剂盒来帮助追踪病毒的传播。
本综述旨在描述和评估目前的逆转录定量聚合酶链反应（RT-qPCR）诊断试剂盒。进一步深入详细介绍了 COVID-19 诊断程序中使用的几个过程，以展示最佳实践。因此，我们讨论了影响 SARS-COV-2 病毒检测的主要因素以及它们如何影响患者的诊断。
这里强调并讨论了可能干扰 RT-PCR 分析的几个因素，例如样本的病毒载量、采集地点、采集方法、样本储存、运输、引物和探针在不同品牌试剂盒中的错配/二聚化。这是一项先驱性研究，旨在讨论可能导致 SARS-CoV-2 的 RT-qPCR 诊断出现错误解释的因素。本研究旨在帮助读者了解 RT-PCR 检测 SARS-CoV-2 出现不良结果背后很可能的原因，以及可以采取哪些措施来获得可靠的诊断。
