國家衛生研究院 NHRI:Item 3990099045/12527
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    題名: nanoMLST: Accurate multilocus sequence typing using Oxford Nanopore Technologies MinION with a dual-barcode approach to multiplex large numbers of samples
    作者: Liou, CH;Wu, HC;Liao, YC;Yang Lauderdale, TL;Huang, IW;Chen, FJ
    貢獻者: National Institute of Infectious Diseases and Vaccinology;Institute of Population Health Sciences
    摘要: Multilocus sequence typing (MLST) is one of the most commonly used methods for studying microbial lineage worldwide. However, the traditional MLST process using Sanger sequencing is time-consuming and expensive. We have designed a workflow that simultaneously sequenced seven full-length housekeeping genes of 96 meticillin-resistant Staphylococcus aureus isolates with dual-barcode multiplexing using just a single flow cell of an Oxford Nanopore Technologies MinION system, and then we performed bioinformatic analysis for strain typing. Fifty-one of the isolates comprising 34 sequence types had been characterized using Sanger sequencing. We demonstrate that the allele assignments obtained by our nanopore workflow (nanoMLST, available at https://github.com/jade-nhri/nanoMLST) were identical to those obtained by Sanger sequencing (359/359, with 100 % agreement rate). In addition, we estimate that our multiplex system is able to perform MLST for up to 1000 samples simultaneously; thus, providing a rapid and cost-effective solution for molecular typing.
    日期: 2020-03
    關聯: Microbial Genomics. 2020 Mar;6(3):Article number 00336.
    Link to: http://dx.doi.org/10.1099/mgen.0.000336
    JIF/Ranking 2023: http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=NHRI&SrcApp=NHRI_IR&KeyISSN=2057-5858&DestApp=IC2JCR
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