hsp65 PCR-restriction analysis (PRA) with capillary electrophoresis for species identification and differentiation of Mycobacterium kansasii and Mycobacterium chelonae-Mycobacterium abscessus group

Int J Infect Dis. 2012 Mar;16(3):e193-7. doi: 10.1016/j.ijid.2011.11.011. Epub 2012 Jan 11.

Abstract

Objectives: The aim of the present study was to identify and differentiate Mycobacterium kansasii and Mycobacterium chelonae-Mycobacterium abscessus group strains isolated from clinical and environmental sources in different countries.

Methods: PCR-restriction analysis of the hsp65 gene (PRA) with automated capillary electrophoresis was applied to the isolates previously identified by conventional biochemical testing and the molecular INNO-LiPA MYCOBACTERIA assay.

Results: PRA performed very well in comparison with the two other methods (96.4% concordance). Among 27 M. kansasii isolates, this method detected five genetic types, of which type 1 represented the most common clinical isolate, as it is worldwide. PRA differentiated 29 M. chelonae-M. abscessus group isolates into Mycobacterium immunogenum type 2 (n=13), M. chelonae (n=12), and M. abscessus types 1 (n=1) and 2 (n=1). M. immunogenum was the most frequent (69%) isolate from humans, but only one of 11 cases was clinically significant. M. chelonae was the most commonly (83%) recovered from water. PRA also identified two isolates with hsp65 alleles representing previously unreported patterns.

Conclusions: PRA based on automated capillary electrophoresis is a rapid, simple, and reliable method for the identification and differentiation of both clinically relevant and environmental isolates of M. kansasii and M. chelonae-M. abscessus group.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Bacterial Typing Techniques / methods*
  • Chaperonin 60 / genetics*
  • Chaperonin 60 / metabolism
  • DNA, Bacterial / genetics*
  • Electrophoresis, Capillary / methods*
  • Genotype
  • Humans
  • Mycobacterium chelonae / classification*
  • Mycobacterium chelonae / genetics
  • Mycobacterium chelonae / isolation & purification
  • Mycobacterium kansasii / classification*
  • Mycobacterium kansasii / genetics
  • Mycobacterium kansasii / isolation & purification
  • Polymerase Chain Reaction / methods
  • Polymorphism, Restriction Fragment Length
  • Sequence Analysis, DNA

Substances

  • Bacterial Proteins
  • Chaperonin 60
  • DNA, Bacterial
  • heat-shock protein 65, Mycobacterium