Quadram Institute Bioscience, Norwich Research Park, Norwich, NR4 7UQ, UK.
Grupo de Investigación Biomédica en Sepsis - BioSepsis. Hospital Universitario Rio Hortega/Instituto de Investigación Biomédica de Salamanca (IBSAL), Valladolid/Salamanca, Spain.
Genome Med. 2021 Feb 9;13(1):21. doi: 10.1186/s13073-021-00839-5.
We present CoronaHiT, a platform and throughput flexible method for sequencing SARS-CoV-2 genomes (≤ 96 on MinION or > 96 on Illumina NextSeq) depending on changing requirements experienced during the pandemic. CoronaHiT uses transposase-based library preparation of ARTIC PCR products. Method performance was demonstrated by sequencing 2 plates containing 95 and 59 SARS-CoV-2 genomes on nanopore and Illumina platforms and comparing to the ARTIC LoCost nanopore method. Of the 154 samples sequenced using all 3 methods, ≥ 90% genome coverage was obtained for 64.3% using ARTIC LoCost, 71.4% using CoronaHiT-ONT and 76.6% using CoronaHiT-Illumina, with almost identical clustering on a maximum likelihood tree. This protocol will aid the rapid expansion of SARS-CoV-2 genome sequencing globally.
我们提出了 CoronaHiT，这是一种平台和通量灵活的方法，可根据大流行期间经验到的变化需求对 SARS-CoV-2 基因组（MinION 上的≤96 或 Illumina NextSeq 上的>96）进行测序。CoronaHiT 使用转座酶为基础的 ARTIC PCR 产物文库制备。通过在纳米孔和 Illumina 平台上对包含 95 和 59 个 SARS-CoV-2 基因组的 2 个板进行测序，并与 ARTIC LoCost 纳米孔方法进行比较，验证了该方法的性能。在使用所有 3 种方法测序的 154 个样本中，使用 ARTIC LoCost 获得了≥90%的基因组覆盖率的样本占 64.3%，使用 CoronaHiT-ONT 的占 71.4%，使用 CoronaHiT-Illumina 的占 76.6%，最大似然树的聚类几乎相同。该方案将有助于在全球范围内快速扩展 SARS-CoV-2 基因组测序。
