Abstract:
:Members of the Mycobacterium chelonae complex (MCC), namely M. chelonae, Mycobacterium abscessus and Mycobacterium immunogenum, have been implicated in nosocomial infections and 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 the existing methods. Here we report a simple and rapid method for unambiguous identification and differentiation of the three-member species of the MCC group with PCR-restriction analysis targeting a 667-bp segment of a variable region of the 65-kDa-heat shock protein (hsp65) gene. This assay, described as Amplified hsp65 Restriction Analysis (AHSPRA), can discriminate all the three individual species using a one-step restriction digestion using either BbvI or Eco0109I. The enzyme NarI can differentiate M. immunogenum from the other two MCC species (M. chelonae and M. abscessus). The developed method was validated using several non-MCC reference species of other rapidly growing mycobacteria (RGM) and MCC field isolates from MWF samples. Direct cell-lysis was used instead of the conventional DNA template preparation, which improved the rapidity, simplicity and adaptability of the developed method. The results suggest that the developed method can unambiguously differentiate species of the M. chelonae complex from other RGM species and from one another.
journal_name
Mol Cell Probesjournal_title
Molecular and cellular probesauthors
Selvaraju SB,Khan IU,Yadav JSdoi
10.1016/j.mcp.2004.09.007subject
Has Abstractpub_date
2005-04-01 00:00:00pages
93-9issue
2eissn
0890-8508issn
1096-1194pii
S0890-8508(04)00095-7journal_volume
19pub_type
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journal_title:Molecular and cellular probes
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doi:10.1016/j.mcp.2006.09.001
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pub_type: 杂志文章,meta分析
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