論文

国際誌
2022年

COVID-19 Whole-Genome Resequencing with Redundant Tiling PCR and Subtract-Based Amplicon Normalization Successfully Characterized SARS-CoV-2 Variants in Clinical Specimens.

Interdisciplinary perspectives on infectious diseases
  • Tatsuki Sugi
  • ,
  • Mizanur Rahman
  • ,
  • Rummana Rahim
  • ,
  • Abu Hasan
  • ,
  • Naoko Kawai
  • ,
  • Kyoko Hayashida
  • ,
  • Junya Yamagishi

2022
開始ページ
2109641
終了ページ
2109641
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1155/2022/2109641

With an increasing number of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) sequences gathered worldwide, we recognize that deletion mutants and nucleotide substitutions that may affect whole-genome sequencing are accumulating. Here, we propose an additional strategy for tiling PCR for whole-genome resequencing, which can make the pipeline robust for mutations at the primer annealing site by a redundant amplicon scheme. We further demonstrated that subtracting overrepresented amplicons from the multiplex PCR products reduced the bias of the next-generation sequencing (NGS) library, resulting in decreasing required sequencing reads per sample. We applied this sequencing strategy to clinical specimens collected in Bangladesh. More than 80% out of the 304 samples were successfully sequenced. Less than 5% were ambiguous nucleotides, and several known variants were detected. With the additional strategies presented here, we believe that whole-genome resequencing of SARS-CoV-2 from clinical samples can be optimized.

リンク情報
DOI
https://doi.org/10.1155/2022/2109641
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/36212105
PubMed Central
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9534710
ID情報
  • DOI : 10.1155/2022/2109641
  • PubMed ID : 36212105
  • PubMed Central 記事ID : PMC9534710

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