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rpoB sequence-based identification of Mycobacterium avium complex species

Toidi Adékambi

Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes, UMR, CNRS-IRD 6236, IFR 48 Faculté de Médecine, Université de la Méditerranée, Marseille, France

Correspondence
Michel Drancourt
Michel.Drancourt{at}medecine.univ-mrs.fr

The Mycobacterium avium complex (MAC) comprises slowly growing mycobacteria responsible for opportunistic infections and zoonoses. The ability to speciate MAC isolates in the clinical microbiology laboratory is critical for determining the organism implicated in clinical disease and for epidemiological investigation of the source of infection. Investigation of a 711 bp variable fragment of rpoB flanked by the Myco-F/Myco-R primers found a 0.7–5.1 % divergence among MAC reference strains, with Mycobacterium chimaera and Mycobacterium intracellulare being the most closely related. Using a 0.7 % divergence cut-off, 83 % of 100 clinical isolates, which had been previously identified by phenotypic characteristics and 16S–23S rDNA intergenic spacer (ITS) probing, were identified as M. avium, 8 % as M. intracellulare and 2 % as M. chimaera. The uniqueness of seven isolates, exhibiting <99.3 % rpoB sequence similarity with MAC reference strains, was confirmed by 16S rDNA, ITS and hsp65 sequencing and phylogenetic analyses. Partial rpoB gene sequencing using the Myco-F/Myco-R primers permits one-step identification of MAC isolates at the species level and the detection of potentially novel MAC species.

Present address: Mycobacteriology Laboratory Branch, Division of Tuberculosis Elimination, Centers for Disease Control and Prevention, Atlanta, GA, USA.

The GenBank/EMBL/DDBJ accession numbers for the Mycobacterium avium complex sequences determined in this study are listed in Table 3.

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