Detection of Mycobacterium immunogenum by real-time quantitative Taqman PCR
| Autor: | Alexandra Fluri, Marco Gerber, Glenn Rhodes, Roger W. Pickup, Alan Henderson, Andrea Ruefenacht |
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| Rok vydání: | 2007 |
| Předmět: |
Microbiology (medical)
Molecular Sequence Data Mycobacterium chelonae Early detection Microbiology Polymerase Chain Reaction law.invention Mycobacterium Bacterial Proteins law TaqMan Humans Molecular Biology Polymerase chain reaction biology Base Sequence DNA-Directed RNA Polymerases biology.organism_classification Molecular biology United Kingdom United States Occupational Diseases genomic DNA Real-time polymerase chain reaction Mycobacterium immunogenum Detection rate Water Microbiology Sequence Alignment Alveolitis Extrinsic Allergic |
| Zdroj: | Journal of microbiological methods. 73(3) |
| ISSN: | 0167-7012 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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