Multiplex amplicon sequencing for the comprehensive genotyping of <i>Mycoplasma pneumoniae</i>.

ABSTRACT: Major genotyping methods used to characterize Mycoplasma pneumoniae strains are based on various experimental approaches that need to be implemented in parallel for each strain. In this study, we developed a comprehensive workflow based on amplicon sequencing using next-generation sequencing. This workflow comprised PCR amplification with seven tubes, collection into single tubes, shotgun sequencing, de novo assembly separating each target into individual contigs, and in silico genotyping. The results for p1, orf6, multilocus sequence types, 23S ribosomal RNA gene mutations conferring macrolide resistance, and single-nucleotide polymorphisms identifying the p1 type 1 lineage were obtained simultaneously. The genotyping accuracy was confirmed by com

SUBMITTER: Kubota H 

PROVIDER: S-EPMC12210885 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature