title: Use of BACTEC radiometric culture method and polymerase chain reaction for the rapid screening of faeces and tissues for Mycobacterium paratuberculosis.
abstract: The BACTEC radiometric culture method for detection of Mycobacterium paratuberculosis was evaluated on faeces from cattle on a farm in quarantine for Johne's disease. A multiplex polymerase chain reaction (PCR) based on the IS900 sequence specific for M paratuberculosis and a genus specific 16S rRNA region was developed and used to test cultures showing evidence of mycobacterial growth in the BACTEC liquid radiometric culture medium. Using the BACTEC-PCR combination, confirmation of M paratuberculosis from faeces and tissue of clinically affected animals was achieved within 2 to 4 weeks and 1 week, respectively, a substantial improvement on traditional culture and identification methods. The PCR provided rapid exclusion of M paratuberculosis when other Mycobacterium spp were grown. The radiometric culture medium proved to be very sensitive for culturing Mycobacterium spp.
authors.affiliation: Animal Health Laboratories, Department of Agriculture, South Perth, Western Australia.
authors.forename: D V
authors.initials: DV
authors.lastname: Cousins
authors.forename: R J
authors.initials: RJ
authors.lastname: Evans
authors.forename: B R
authors.initials: BR
authors.lastname: Francis
mesh_terms: D000818:Animals; D001315:Australia / Q000453:epidemiology; D001483:Base Sequence; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000453:epidemiology / Q000517:prevention & control*; D002432:Cecum / Q000382:microbiology*; D017931:DNA Primers / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D005243:Feces / Q000382:microbiology*; D007082:Ileum / Q000382:microbiology*; D008198:Lymph Nodes / Q000382:microbiology*; D008403:Mass Screening / Q000379:methods / Q000662:veterinary*; D008969:Molecular Sequence Data; D016927:Mycobacterium avium subsp. paratuberculosis / Q000235:genetics / Q000302:isolation & purification*; D010283:Paratuberculosis / Q000175:diagnosis / Q000453:epidemiology / Q000517:prevention & control*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011874:Radiometry / Q000379:methods / Q000662:veterinary*
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial