Detection of viable Mycobacterium avium subsp. paratuberculosis using luciferase reporter systems

Foodborne Pathog Dis. Winter 2004;1(4):258-66. doi: 10.1089/fpd.2004.1.258.

Abstract

Plasmid- and phage-based firefly luciferase reporter constructs were evaluated as rapid detection systems for viable Mycobacterium avium subsp. paratuberculosis (MAP). A MAP strain bearing a luciferase-encoding plasmid was detectable at 100 cells/mL in skim milk and 1000 cells/mL in whole milk. Three luciferase-encoding mycobacteriophage were evaluated for detection of wild-type MAP. The best of these, phAE85, allowed detection of >1000 cells/mL within 24-48 h. Membrane filtration did not improve the sensitivity of detection for either plasmid or phage reporters. Luciferase reporters show promise for rapid detection of viable MAP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Colony Count, Microbial
  • Food Contamination / analysis*
  • Food Microbiology
  • Luciferases, Bacterial / genetics*
  • Luciferases, Bacterial / metabolism
  • Milk / microbiology*
  • Mycobacterium avium subsp. paratuberculosis / isolation & purification*
  • Plasmids
  • Time Factors

Substances

  • Luciferases, Bacterial