Journal
JOURNAL OF MICROBIOLOGICAL METHODS
Volume 73, Issue 3, Pages 266-268Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.mimet.2008.03.006
Keywords
Mycobacterium immunogenum; Mycobacterium chelonae complex; hypersensitivity pneumonitis; metal working fluids; real-time PCR; Taqman
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Funding
- Natural Environment Research Council [CEH010021] Funding Source: researchfish
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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 3.4 x 10(1) and 1.9 x 10(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. (C) 2008 Elsevier B.V. All rights reserved.
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