商业双靶标 qRT-PCR 诊断检测中 SARS-CoV-2 ORF1ab 扩增减弱。
Diminished amplification of SARS-CoV-2 ORF1ab in a commercial dual-target qRT-PCR diagnostic assay.
机构信息
Department of Viroscience, Erasmus MC, Rotterdam, the Netherlands.
Department of Viroscience, Erasmus MC, Rotterdam, the Netherlands.
出版信息
J Virol Methods. 2022 Feb;300:114397. doi: 10.1016/j.jviromet.2021.114397. Epub 2021 Dec 1.
Here we describe a SARS-CoV-2 variant with diminished amplification of the ORF ORF1ab target in the Cobas® dual-target SARS-CoV-2 assay resulting in a discrepancy of Ct-values (Ct-value 20.7 for the E-gene and Ct-value 30.2 for ORF1ab). Five unique nucleotide mutations were identified in ORF1ab: C11450A (nsp10) C14178T (RdRp), G15006T (RdRp), G18394T (Hel), and G20995T (Hel). This case highlights the importance of surveillance of genomic regions used in molecular diagnostics and the importance of the public release of target regions used to update commercial and in-house developed SARS-CoV-2 PCR tests. This work underpins the importance of using dual-targets in molecular diagnostic assays to limit the change of false-negative results due to primer and/or probe mismatches.
在这里，我们描述了一种 SARS-CoV-2 变异株，其 ORF1ab 目标在 Cobas®双靶 SARS-CoV-2 检测中的扩增能力减弱，导致 Ct 值出现差异（E 基因的 Ct 值为 20.7，而 ORF1ab 的 Ct 值为 30.2）。在 ORF1ab 中发现了 5 个独特的核苷酸突变：C11450A（nsp10）C14178T（RdRp）、G15006T（RdRp）、G18394T（Hel）和 G20995T（Hel）。该病例强调了监测分子诊断中使用的基因组区域以及发布用于更新商业和内部开发的 SARS-CoV-2 PCR 测试的目标区域的重要性。这项工作强调了在分子诊断检测中使用双靶标来限制由于引物和/或探针不匹配而导致假阴性结果的变化的重要性。
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