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The rapidly growing mycobacteriumM. abscessus sensu lato is the causative agent of emerging pulmonary and skin diseases and of infections following cosmetic surgery and postsurgical procedures.M. abscessus sensu latocan be divided into at least three subspecies:M. abscessus subsp.abscessus ,M. abscessus subsp.massiliense , andM. abscessus subsp.bolletii . Clinical isolates of rapidly growing mycobacteria were previously identified asM. abscessus by DNA-DNA hybridization. More than 30% of these 117 clinical isolates were differentiated asM. abscessus subsp.massiliense using combinations of multilocus genotyping analyses. A much more cost-effective technique to distinguishM. abscessus subsp.massiliense fromM. abscessus subsp.abscessus , a multiplex PCR assay, was developed using the whole-genome sequence ofM. abscessus subsp.massiliense JCM15300 as a reference. Several primer sets were designed for single PCR to discriminate between the strains based on amplicons of different sizes. Two of these single-PCR target sites were chosen for development of the multiplex PCR assay. Multiplex PCR was successful in distinguishing clinical isolates ofM. abscessus subsp.massiliense from samples previously identified asM. abscessus . This approach, which spans whole-genome sequencing and clinical diagnosis, will facilitate the acquisition of more-precise information about bacterial genomes, aid in the choice of more relevant therapies, and promote the advancement of novel discrimination and differential diagnostic assays.

Discrimination of Mycobacterium abscessus subsp. massiliense from Mycobacterium abscessus subsp. abscessus in Clinical Isolates by Multiplex PCR