Department of Microbiology, Infectious Diseases and Immunology, Université de Montréal, Montréal, Québec, Canada.
Immunopathology Axis, Centre de Recherche du Centre Hospitalier de l'Université de Montréal, Montréal, Québec, Canada.
Diagn Microbiol Infect Dis. 2021 Dec;101(4):115521. doi: 10.1016/j.diagmicrobio.2021.115521. Epub 2021 Aug 21.
SARS-CoV-2 whole genome sequencing is a molecular biology tool performed to support many aspects of the response to the pandemic. Freezing of primary clinical nasopharyngeal swabs and shipment to reference laboratories is usually required for sequencing. Cobas PCR Media transport medium facilitates high throughput SARS-CoV-2 RT-PCR analyses on cobas platforms. The manufacturer doesn't recommend freezing this transport medium because of risks of degrading molecular templates and impairing test results. Our objective was to compare the quality and results of SARS-CoV-2 genomic sequencing when performed on fresh or frozen samples in cobas PCR Media. Viral genome sequencing was performed using Oxford Nanopore Technologies MinION platform. Sequencing performance, quality and results did not significantly differ between fresh and frozen samples (n = 10). Freezing of cobas PCR Media does not negatively affect SARS-CoV-2 RNA sequencing results and it is therefore a suitable transport medium for outsourcing sequencing analyses to reference laboratories.
SARS-CoV-2 全基因组测序是一种分子生物学工具,用于支持应对大流行的许多方面。通常需要将原始临床鼻咽拭子冷冻并运送到参考实验室进行测序。Cobas PCR 介质运输培养基有助于在 cobas 平台上进行高通量 SARS-CoV-2 RT-PCR 分析。由于存在降解分子模板和影响测试结果的风险,制造商不建议冷冻该运输培养基。我们的目的是比较在 cobas PCR 介质中对新鲜或冷冻样本进行 SARS-CoV-2 基因组测序时的质量和结果。使用牛津纳米孔技术 MinION 平台进行病毒基因组测序。新鲜和冷冻样本之间的测序性能、质量和结果没有显著差异(n=10)。冷冻 cobas PCR 介质不会对 SARS-CoV-2 RNA 测序结果产生负面影响,因此它是一种适合将测序分析外包给参考实验室的运输培养基。