Detection of Mycobacterium immunogenum by real-time quantitative Taqman PCR.
A quantitative real-time 5'-nuclease (Taqman) PCR technique was developed to specifically detect Mycobacterium immunogenum. rpoB-specific primers and Taqman probe were evaluated for detection of M. immunogenum DNA extracted from pure cultures and from industrial metal working fluids (MWFs). Specificity was confirmed and the sensitivity of detection of M. immunogenum genomic DNA was shown to be approximately 9 fg (2 cell equivalents). When tested on industrial metal working fluids from the UK and USA from which no M. immunogenum CFU were recovered, the assay detected between 3.4x10(1) and 1.9x10(4) cell equivalents (CE) per ml, and increased the detection rate over culture to 37.5% (12 of 32 samples). This assay provides a specific, sensitive and rapid method for the detection of M. immunogenum and is applicable within industry for the early detection of this human pathogen and to the possible prevention of hypersensitivity pneumonitis (HP) in workers.
Rhodes
Glenn
G
Centre for Ecology and Hydrology, Lancaster Environment Centre, Library Avenue, Bailrigg, Lancaster. LA1 4AP, United Kingdom. glenn@ceh.ac.uk
Fluri
Alexandra
A
Gerber
Marco
M
Henderson
Alan
A
Ruefenacht
Andrea
A
Pickup
Roger W
RW
D000542:Alveolitis, Extrinsic Allergic / Q000517:prevention & control*; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000235:genetics / Q000302:isolation & purification*; D009784:Occupational Diseases / Q000517:prevention & control*; D016133:Polymerase Chain Reaction / Q000379:methods*; D016415:Sequence Alignment; D006113:United Kingdom; D014481:United States; D014871:Water Microbiology
D001426:Bacterial Proteins; D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit