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

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作者
Dave J. Baker
Alp Aydin
Thanh Le-Viet
Gemma L. Kay
Steven Rudder
Leonardo de Oliveira Martins
Ana P. Tedim
Anastasia Kolyva
Maria Diaz
Nabil-Fareed Alikhan
Lizzie Meadows
Andrew Bell
Ana Victoria Gutierrez
Alexander J. Trotter
Nicholas M. Thomson
Rachel Gilroy
Luke Griffith
Evelien M. Adriaenssens
Rachael Stanley
Ian G. Charles
Ngozi Elumogo
John Wain
Reenesh Prakash
Emma Meader
Alison E. Mather
Mark A. Webber
Samir Dervisevic
Andrew J. Page
Justin O’Grady
机构
[1] Quadram Institute Bioscience,
[2] Norwich Research Park,undefined
[3] Grupo de Investigación Biomédica en Sepsis - BioSepsis. Hospital Universitario Rio Hortega/Instituto de Investigación Biomédica de Salamanca (IBSAL),undefined
[4] Norfolk and Norwich University Hospital,undefined
[5] University of East Anglia,undefined
[6] Norwich Research Park,undefined
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关键词
SARS-CoV-2; Nanopore; Sequencing; NGS; Genome; Genetic; Multiplexing; ARTIC;
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摘要
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.
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