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Detection of Mycobacterium immunogenum by real-time quantitative Taqman PCR

Research output: Contribution to Journal/MagazineJournal articlepeer-review

Published
  • Glenn Rhodes
  • Alexandra Fluri
  • Marco Gerber
  • Alan Henderson
  • Andrea Ruefenacht
  • Roger W. Pickup
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<mark>Journal publication date</mark>30/06/2008
<mark>Journal</mark>Journal of Microbiological Methods
Issue number3
Volume73
Number of pages3
Pages (from-to)266-268
Publication StatusPublished
<mark>Original language</mark>English

Abstract

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 × 101 and 1.9 × 104 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.