RRC ID 34075
Author Nakanaga K, Sekizuka T, Fukano H, Sakakibara Y, Takeuchi F, Wada S, Ishii N, Makino M, Kuroda M, Hoshino Y.
Title Discrimination of Mycobacterium abscessus subsp. massiliense from Mycobacterium abscessus subsp. abscessus in clinical isolates by multiplex PCR.
Journal J. Clin. Microbiol.
Abstract The rapidly growing mycobacterium M. 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 lato can be divided into at least three subspecies: M. abscessus subsp. abscessus, M. abscessus subsp. massiliense, and M. abscessus subsp. bolletii. Clinical isolates of rapidly growing mycobacteria were previously identified as M. abscessus by DNA-DNA hybridization. More than 30% of these 117 clinical isolates were differentiated as M. abscessus subsp. massiliense using combinations of multilocus genotyping analyses. A much more cost-effective technique to distinguish M. abscessus subsp. massiliense from M. abscessus subsp. abscessus, a multiplex PCR assay, was developed using the whole-genome sequence of M. 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 of M. abscessus subsp. massiliense from samples previously identified as M. 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.
Volume 52(1)
Pages 251-9
Published 2014-1
DOI 10.1128/JCM.01327-13
PII JCM.01327-13
PMID 24197885
PMC PMC3911466
MeSH Bacteriological Techniques / methods* DNA Primers / genetics DNA, Bacterial / chemistry DNA, Bacterial / genetics DNA, Ribosomal / chemistry DNA, Ribosomal / genetics Genome, Bacterial Humans Molecular Diagnostic Techniques / methods* Molecular Sequence Data Multiplex Polymerase Chain Reaction / methods* Mycobacterium / classification* Mycobacterium / genetics Mycobacterium / isolation & purification* Mycobacterium Infections, Nontuberculous / microbiology* RNA, Ribosomal, 16S / genetics Respiratory Tract Infections / microbiology Sequence Analysis, DNA Skin Diseases, Bacterial / microbiology
IF 4.054
Times Cited 11
General Microbes JCM 6388 JCM 13569 JCM 15297 JCM 15300