This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Stanley, E. C.
Right arrow Articles by Rees, C. E. D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Stanley, E. C.
Right arrow Articles by Rees, C. E. D.
Agricola
Right arrow Articles by Stanley, E. C.
Right arrow Articles by Rees, C. E. D.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, March 2007, p. 1851-1857, Vol. 73, No. 6
0099-2240/07/$08.00+0     doi:10.1128/AEM.01722-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Development of a New, Combined Rapid Method Using Phage and PCR for Detection and Identification of Viable Mycobacterium paratuberculosis Bacteria within 48 Hours{triangledown}

Emma C. Stanley,1 Richard J. Mole,2 Rebecca J. Smith,3 Sarah M. Glenn,3 Michael R. Barer,3 Michael McGowan,4 and Catherine E. D. Rees1*

Division of Food Sciences, School of Biosciences, Sutton Bonington Campus, University of Nottingham, Sutton Bonington LE12 5RD, United Kingdom,1 Biotec Laboratories, 32 Anson Road, Marltesham Heath, Ipswich IP5 3RG, United Kingdom,2 Department of Infection, Immunity and Inflammation, University of Leicester, Medical Sciences Building, PO Box 138, University Rd., Leicester LE1 9HN, United Kingdom,3 The University of Queensland, School of Veterinary Science, Brisbane, Queensland, Australia4

Received 21 July 2006/ Accepted 28 December 2006

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.

* Corresponding author. Mailing address: Division of Food Sciences, School of Biosciences, Sutton Bonington Campus, University of Nottingham, Sutton Bonington LE12 5RD, United Kingdom. Phone: 44 115 9516167. Fax: 44 115 9516162. E-mail: cath.rees{at}nottingham.ac.uk.

{triangledown} Published ahead of print on 26 January 2007.

Applied and Environmental Microbiology, March 2007, p. 1851-1857, Vol. 73, No. 6
0099-2240/07/$08.00+0     doi:10.1128/AEM.01722-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Foddai, A., Elliott, C. T., Grant, I. R. (2009). Optimization of a Phage Amplification Assay To Permit Accurate Enumeration of Viable Mycobacterium avium subsp. paratuberculosis Cells. Appl. Environ. Microbiol. 75: 3896-3902 [Abstract] [Full Text]  
  • Altic, L. C., Rowe, M. T., Grant, I. R. (2007). UV Light Inactivation of Mycobacterium avium subsp. paratuberculosis in Milk as Assessed by FASTPlaqueTB Phage Assay and Culture. Appl. Environ. Microbiol. 73: 3728-3733 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»