nanoMLST: accurate multilocus sequence typing using Oxford Nanopore Technologies MinION with a dual-barcode approach to multiplex large numbers of samples

• DOI: 10.1099/mgen.0.000336
• Publication Date: 2020-02-17T16:13:40Z
• Authors: Ci-Hong Liou; Han-Chieh Wu; Yu-Chieh Liao; Tsai-Ling Yang La...; I-Wen Huang; Feng-Jui Chen
• Keywords: Minion; Barcode; Multiplex; Multilocus sequence typing; Sequence (biology)

Multilocus sequence typing (MLST) is one of the most commonly used methods for studying microbial lineage worldwide. However, traditional MLST process using Sanger sequencing time-consuming and expensive. We have designed a workflow that simultaneously sequenced seven full-length housekeeping genes 96 meticillin-resistant Staphylococcus aureus isolates with dual-barcode multiplexing just single flow cell an Oxford Nanopore Technologies MinION system, then we performed bioinformatic analysis strain typing. Fifty-one comprising 34 types had been characterized sequencing. demonstrate allele assignments obtained by our nanopore (nanoMLST, available at https://github.com/jade-nhri/nanoMLST) were identical to those (359/359, 100 % agreement rate). In addition, estimate multiplex system able perform up 1000 samples simultaneously; thus, providing rapid cost-effective solution molecular