Use of BACTEC radiometric culture method and polymerase chain reaction for the rapid screening of faeces and tissues for Mycobacterium paratuberculosis

Aust Vet J. 1995 Dec;72(12):458-62. doi: 10.1111/j.1751-0813.1995.tb03489.x.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Australia / epidemiology
  • Base Sequence
  • Cattle
  • Cattle Diseases / diagnosis
  • Cattle Diseases / epidemiology
  • Cattle Diseases / prevention & control*
  • Cecum / microbiology*
  • DNA Primers / analysis
  • DNA Primers / chemistry
  • DNA Primers / genetics
  • DNA, Bacterial / analysis
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Feces / microbiology*
  • Ileum / microbiology*
  • Lymph Nodes / microbiology*
  • Mass Screening / methods
  • Mass Screening / veterinary*
  • Molecular Sequence Data
  • Mycobacterium avium subsp. paratuberculosis / genetics
  • Mycobacterium avium subsp. paratuberculosis / isolation & purification*
  • Paratuberculosis / diagnosis
  • Paratuberculosis / epidemiology
  • Paratuberculosis / prevention & control*
  • Polymerase Chain Reaction / methods
  • Polymerase Chain Reaction / veterinary*
  • Radiometry / methods
  • Radiometry / veterinary*

Substances

  • DNA Primers
  • DNA, Bacterial