國家衛生研究院 NHRI:Item 3990099045/14212

NHRI > National Institute of Infectious Diseases and Vaccinology > Feng-Jui Chen > Periodical Articles > Item 3990099045/14212

Please use this identifier to cite or link to this item: http://ir.nhri.org.tw/handle/3990099045/14212

Title:	Rapid and routine molecular typing using multiplex polymerase chain reaction and minION sequencer
Authors:	Liao, YC;Wu, HC;Liou, CH;Lauderdale, TLY;Huang, IW;Lai, JF;Chen, FJ
Contributors:	National Institute of Infectious Diseases and Vaccinology;Institute of Population Health Sciences
Abstract:	Molecular typing is an essential tool that has been extensively applied in laboratories as well as in clinical settings. Next-generation sequencing technologies promise high-throughput and cost-effective molecular applications; however, the accessibility of these technologies is limited due to the high capital cost. Oxford Nanopore Technologies (ONT) offers a MinION device with the advantages of real-time data analysis, rapid library preparation, and low cost per test. However, the advantages of the MinION device are often overshadowed by its lower raw accuracy. Herein, we present a concise multilocus sequence typing protocol of Staphylococcus aureus using multiplex polymerase chain reaction and Rapid Barcoding Kit for barcoding and MinION device for sequencing. Moreover, to clarify the effects of carryover DNA on tasks that require high sequence accuracy, we used the MinION flow cell in successive runs of washing and reusing. Our results revealed that the MinION flow cell could achieve accurate typing of a total of 467 samples with 3,269 kilobase-long genes within a total of 5 runs. This thus demonstrates the effectiveness of a portable nanopore MinION sequencer in providing accurate, rapid, and routine molecular typing.
Date:	2022-03-29
Relation:	Frontiers in Microbiology. 2022 Mar 29;13:Article number 875347.
Link to:	http://dx.doi.org/10.3389/fmicb.2022.875347
JIF/Ranking 2023:	http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=NHRI&SrcApp=NHRI_IR&KeyISSN=1664-302X&DestApp=IC2JCR
Cited Times(WOS):	https://www.webofscience.com/wos/woscc/full-record/WOS:000790452200001
Cited Times(Scopus):	https://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=85128468757
Appears in Collections:	[Feng-Jui Chen] Periodical Articles [Tsai-Ling Yang Lauderdale] Periodical Articles [Yu-Chieh Liao] Periodical Articles

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