CoronaHiT: high-throughput sequencing of SARS-CoV-2 genomes.

CoronaHiT: high-throughput sequencing of SARS-CoV-2 genomes.
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DOI:
10.1186/s13073-021-00839-5
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发表时间:
2021-02-09
期刊:
影响因子:
12.3
通讯作者:
O'Grady J
O'Grady J
中科院分区:
生物学1区
文献类型:
--
作者:
Baker DJ;Aydin A;Le-Viet T;Kay GL;Rudder S;de Oliveira Martins L;Tedim AP;Kolyva A;Diaz M;Alikhan NF;Meadows L;Bell A;Gutierrez AV;Trotter AJ;Thomson NM;Gilroy R;Griffith L;Adriaenssens EM;Stanley R;Charles IG;Elumogo N;Wain J;Prakash R;Meader E;Mather AE;Webber MA;Dervisevic S;Page AJ;O'Grady J

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我们提出了CoronaHiT,这是一种平台和通量灵活的方法,用于对SARS-CoV-2基因组进行测序(在MinION上≤ 96或在Illumina NextSeq上> 96),这取决于大流行期间不断变化的要求。CoronaHiT使用ARTIC PCR产物的基于转座酶的文库制备。通过在纳米孔和Illumina平台上对含有95个和59个SARS-CoV-2基因组的2个平板进行测序并与ARTIC LoCost纳米孔方法进行比较来证明方法性能。在使用所有3种方法测序的154个样本中,使用ARTIC LoCost获得≥ 90%的基因组覆盖率,使用CoronaHiT-ONT获得71.4%,使用CoronaHiT-Illumina获得76.6%,在最大似然树上具有几乎相同的聚类。该方案将有助于SARS-CoV-2基因组测序在全球范围内的快速扩展。在线版本包含补充材料,可通过10.1186/s13073-021-00839-5获得。
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. The online version contains supplementary material available at 10.1186/s13073-021-00839-5.
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