Comparison of three DNA extraction methods for molecular confirmation of Mycobacterium avium subspecies paratuberculosis from the VersaTrek™ liquid cultures of bovine fecal samples

J Microbiol Methods. 2018 Sep:152:27-30. doi: 10.1016/j.mimet.2018.07.012. Epub 2018 Jul 19.

Abstract

We evaluated three DNA extraction methods for confirmation of Mycobacterium avium subspecies paratuberculosis from liquid cultures of bovine feces. Use of DNA Extract All Reagents Kit™ resulted in efficient extraction of amplifiable DNA from higher proportion (96.29%) of known positive samples compared to Chelex-100 resin (25.92%) and polyethylene glycol (0%).

Keywords: DNA extraction; Johne's disease; Liquid culture; Mycobacterium; Paratuberculosis.

MeSH terms

  • Animals
  • Bacteriological Techniques / methods*
  • Bacteriological Techniques / veterinary*
  • Cattle
  • Cattle Diseases / diagnosis
  • Cattle Diseases / microbiology
  • Culture Media / chemistry
  • DNA / chemistry*
  • DNA / isolation & purification
  • Diagnostic Tests, Routine / methods
  • Diagnostic Tests, Routine / veterinary
  • Feces / microbiology*
  • Molecular Conformation*
  • Mycobacterium avium subsp. paratuberculosis / genetics*
  • Mycobacterium avium subsp. paratuberculosis / isolation & purification
  • Mycobacterium avium subsp. paratuberculosis / pathogenicity
  • Paratuberculosis / diagnosis
  • Paratuberculosis / microbiology
  • Sensitivity and Specificity

Substances

  • Culture Media
  • DNA