A new method for species identification and differentiation of Mycobacterium chelonae complex based on amplified hsp65 restriction analysis (AHSPRA)

摘要: Members of the Mycobacterium chelonae complex (MCC), namely M. chelonae, abscessus and immunogenum, have been implicated in nosocomial infections occupational respiratory illnesses like hypersensitivity pneumonitis (HP) associated with contaminated metalworking fluid (MWF) exposures. Close relationship among these member species makes their differentiation cumbersome using existing methods. Here we report a simple rapid method for unambiguous identification three-member MCC group PCR-restriction analysis targeting 667-bp segment variable region 65-kDa-heat shock protein (hsp65) gene. This assay, described as Amplified hsp65 Restriction Analysis (AHSPRA), can discriminate all three individual one-step restriction digestion either BbvI or Eco0109I. The enzyme NarI differentiate immunogenum from other two (M. abscessus). developed was validated several non-MCC reference rapidly growing mycobacteria (RGM) field isolates MWF samples. Direct cell-lysis used instead conventional DNA template preparation, which improved rapidity, simplicity adaptability method. results suggest that unambiguously RGM one another.