Detection of Mycobacterium avium ssp. paratuberculosis in ileocaecal lymph nodes collected from elderly slaughter cows using a semi-nested IS900 polymerase chain reaction

Vet Microbiol. 2011 Dec 29;154(1-2):197-201. doi: 10.1016/j.vetmic.2011.06.033. Epub 2011 Jul 2.

Abstract

The aim of this study was to investigate the occurrence of subclinical Mycobacterium avium spp. paratuberculosis (MAP) infections at slaughter by testing ileocaecal lymph nodes with a semi-nested IS900 PCR. Tissue samples were available within the framework of a parallel study investigating BSE-susceptibility factors in members of BSE-cohorts in the German Federal State of Lower Saxony. Ileocaecal lymph nodes were collected over a 2-year sampling period from 99 slaughter cattle of a mean age of 6.5 years (5.5-7.5 years). A recently developed IS900 semi-nested polymerase chain reaction (snPCR) assay offering a sensitivity of 1 genome equivalent was used for the detection of MAP-DNA. Based on this snPCR, 17 out of the 99 samples gave positive results, indicating a MAP occurrence of 17.17% in the random sample. All PCR products were sequenced for screening of polymorphisms. Nucleotide homologies of 98.5-100% were found with respect to the MAP K10 reference sequence IS900 (GenBank: AE16958). PCR analysis of ileocaecal lymph nodes collected from slaughter cattle proved to be a suitable technique to determine MAP occurrence in the local cattle population.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle / microbiology*
  • Cattle Diseases / diagnosis*
  • Cattle Diseases / epidemiology
  • Cattle Diseases / microbiology
  • DNA, Bacterial / genetics
  • Germany / epidemiology
  • Lymph Nodes / microbiology*
  • Mycobacterium avium subsp. paratuberculosis / genetics
  • Mycobacterium avium subsp. paratuberculosis / isolation & purification*
  • Paratuberculosis / diagnosis*
  • Paratuberculosis / epidemiology
  • Paratuberculosis / microbiology
  • Polymerase Chain Reaction / methods
  • Polymerase Chain Reaction / veterinary
  • Prevalence
  • Sensitivity and Specificity

Substances

  • DNA, Bacterial