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Appl. Environ. Microbiol. doi:10.1128/AEM.01722-06
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Development of a New Rapid Combined Phage and PCR method for the detection and identification of viable Mycobacterium paratuberculosis within 48 h

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

^* To whom correspondence should be addressed. Email: cath.rees{at}nottingham.ac.uk.

	Abstract

The FASTPlaqueTB assay is an established diagnostic aid for the rapid detection of M. tuberculosis from human sputum samples. Using the FASTPlaqueTB assay (FPTB) reagents viable M. avium subspecies paratuberculosis cells were detected in just 24 h as phage plaques. The bacteriophage used does not infect MAP alone, so to add specificity to this assay a PCR-based identification method was introduced to amplify MAP-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 MAP 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 MAP in milk samples in just 48 h.

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• 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]