UMR VITROME, Aix Marseille University, IRD, AP-HM, SSA, IHU-Méditerranée Infection, 13005 Marseille, France.
Service de Biologie, Unité de Microbiologie, Hôpital Mignot, Centre Hospitalier de Versailles, 177 rue de Versailles, 78150 Le Chesnay, France.
Genes (Basel). 2023 Mar 9;14(3):681. doi: 10.3390/genes14030681.
The emergence of the Coronavirus Disease 2019 (COVID-19) pandemic has fostered the use of high-throughput techniques to sequence the entire severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome and track its evolution. The present study proposes a rapid and relatively less expensive sequencing protocol for 384 samples by adapting the use of an Illumina NovaSeq library to an Illumina MiSeq flow cell instrument. The SARS-CoV-2 genome sequences obtained with Illumina NovaSeq and those obtained using MiSeq instruments were compared with the objective to validate the new, modified protocol. A total of 356 (94.6%) samples yielded interpretable sequences using the modified Illumina COVIDSeq protocol, with an average coverage of 91.6%. By comparison, 357 (94.9%) samples yielded interpretable sequences with the standard COVIDSeq protocol, with an average coverage of 95.6%. Our modified COVIDSeq protocol could save 14,155 euros per run and yield results from 384 samples in 53.5 h, compared to four times 55.5 h with the standard Illumina MiSeq protocol. The modified COVIDSeq protocol thus provides high quality results comparable to those obtained with the standard COVIDSeq protocol, four times faster, while saving money.
2019 年冠状病毒病(COVID-19)大流行的出现促进了高通量技术的使用,以对整个严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)基因组进行测序并跟踪其进化。本研究提出了一种快速且相对较便宜的测序方案,用于对 384 个样本进行测序,该方案通过将 Illumina NovaSeq 文库适配到 Illumina MiSeq 流动池仪器上进行使用。使用 Illumina NovaSeq 获得的 SARS-CoV-2 基因组序列与使用 MiSeq 仪器获得的序列进行了比较,目的是验证新的改良方案。使用改良的 Illumina COVIDSeq 方案,有 356 个(94.6%)样本产生了可解释的序列,平均覆盖率为 91.6%。相比之下,有 357 个(94.9%)样本使用标准 COVIDSeq 方案产生了可解释的序列,平均覆盖率为 95.6%。与标准 Illumina MiSeq 方案相比,我们的改良 COVIDSeq 方案每次运行可节省 14,155 欧元,并且可以在 53.5 小时内处理 384 个样本,而标准方案则需要 4 次 55.5 小时。因此,改良的 COVIDSeq 方案可提供与标准 COVIDSeq 方案相当的高质量结果,速度快四倍,同时还可以节省资金。