Development of a new, combined rapid method using phage and PCR for detection and identification of viable Mycobacterium paratuberculosis bacteria within 48 hours

Stanley, Emma C., Mole, Richard J., Smith, Rebecca J., Glenn, Sarah M., Barer, Michael R., McGowan, Michael and Rees, Catherine E. D. (2007) Development of a new, combined rapid method using phage and PCR for detection and identification of viable Mycobacterium paratuberculosis bacteria within 48 hours. Applied and Environmental Microbiology, 73 6: 1851-1857. doi:10.1128/AEM.01722-06

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Author Stanley, Emma C.
Mole, Richard J.
Smith, Rebecca J.
Glenn, Sarah M.
Barer, Michael R.
McGowan, Michael
Rees, Catherine E. D.
Title Development of a new, combined rapid method using phage and PCR for detection and identification of viable Mycobacterium paratuberculosis bacteria within 48 hours
Journal name Applied and Environmental Microbiology   Check publisher's open access policy
ISSN 0099-2240
1098-5336
Publication date 2007-03
Sub-type Article (original research)
DOI 10.1128/AEM.01722-06
Open Access Status File (Publisher version)
Volume 73
Issue 6
Start page 1851
End page 1857
Total pages 7
Place of publication Washington, D. C.
Publisher American Society of Microbiology
Collection year 2008
Language eng
Subject C1
630104 Dairy cattle
300507 Microbiology (excl. Virology)
Abstract The FASTPlaqueTB assay is an established diagnostic aid for the rapid detection of Mycobacterium tuberculosis from human sputum samples. Using the FASTPlaqueTB assay reagents, viable Mycobacterium avium subsp. paratuberculosis cells were detected as phage plaques in just 24 h. The bacteriophage used does not infect M. avium subsp. paratuberculosis alone, so to add specificity to this assay, a PCR-based identification method was introduced to amplify M. avium subsp. paratuberculosis-specific sequences from the DNA of the mycobacterial cell detected by the phage. To give further diagnostic information, a multiplex PCR method was developed to allow simultaneous amplification of either M. avium subsp. paratuberculosis or M. tuberculosis complex-specific sequences from plaque samples. Combining the plaque PCR technique with the phage-based detection assay allowed the rapid and specific detection of viable M. avium subsp. paratuberculosis in milk samples in just 48 h.
Keyword Biotechnology & Applied Microbiology
Microbiology
Avium Subsp Paratuberculosis
Polymerase-chain-reaction
Pulmonary Tuberculosis
Johnes-disease
Crohns-disease
Multiplex-pcr
Culture
Bovis
Differentiation
Streptomycin
Q-Index Code C1
Q-Index Status Confirmed Code

 
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Created: Mon, 18 Feb 2008, 16:42:42 EST