PDF(Pubmed)
Abstract：
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; initially named 2019-nCoV) is responsible for the recent coronavirus disease (COVID-19) pandemic, and polymerase chain reaction (PCR) is the current standard method for diagnosis from patient samples. As PCR assays are prone to sequence mismatches due to mutations in the viral genome, it is important to verify the genomic variability at primer/probe binding regions periodically. This step-by-step protocol describes a bioinformatics approach for an extensive evaluation of the sequence variability within the primer/probe target regions of the SARS-CoV-2 genome. The protocol can be applied to any molecular diagnostic assay of choice using freely available software programs and the ready-to-use multiple sequence alignment (MSA) file provided. Graphic abstract Overview of the sequence tracing protocol. The figure was created using the Library of Science and Medical Illustrations from somersault18:24 licensed under a CC BY-NC-SA 4.0 license (https://creativecommons.org/licenses/by-nc-sa/4.0/). Video abstract: https://youtu.be/M1lV1liWE9k.
摘要：
严重急性呼吸道综合症冠状病毒2（SARS-CoV-2；最初命名为2019-nCoV）是最近冠状病毒病（COVID-19）大流行的原因，聚合酶链反应（PCR）是目前从患者样本中诊断的标准方法。由于PCR分析容易由于病毒基因组中的突变而导致序列错配，定期验证引物/探针结合区域的基因组变异性是重要的。该分步方案描述了一种生物信息学方法，用于广泛评估SARS-CoV-2基因组的引物/探针靶区域内的序列变异性。使用免费可用的软件程序和提供的即用型多序列比对（MSA）文件，该方案可应用于选择的任何分子诊断测定。序列跟踪协议概述。该图是使用由CCBY-NC-SA4.0许可证（https://creativecommons.org/licenses/by-nc-sa/4.0/）许可的somensault18:24科学和医学插图图书馆创建的。视频摘要:https://youtu.是/M1lV1liWE9k。
