Detection of Mycoplasma mycoides subsp. mycoides SC in bronchoalveolar lavage fluids of cows based on a TaqMan real-time PCR discriminating wild type strains from an lppQ(-) mutant vaccine strain used for DIVA-strategies

J Microbiol Methods. 2010 Jun;81(3):211-8. doi: 10.1016/j.mimet.2010.03.025. Epub 2010 Apr 8.

Abstract

Contagious bovine pleuropneumonia (CBPP) is the most serious cattle disease in Africa, caused by Mycoplasma mycoides subsp. mycoides small-colony type (SC). CBPP control strategies currently rely on vaccination with a vaccine based on live attenuated strains of the organism. Recently, an lppQ(-) mutant of the existing vaccine strain T1/44 has been developed (Janis et al., 2008). This T1lppQ(-) mutant strain is devoid of lipoprotein LppQ, a potential virulence attribute of M. mycoides subsp. mycoides SC. It is designated as a potential live DIVA (Differentiating Infected from Vaccinated Animals) vaccine strain allowing both serological and etiological differentiation. The present paper reports on the validation of a control strategy for CBPP in cattle, whereby a TaqMan real-time PCR based on the lppQ gene has been developed for the direct detection of M. mycoides subsp. mycoides SC in ex vivo bronchoalveolar lavage fluids of cows and for the discrimination of wild type strains from the lppQ(-) mutant vaccine strain.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Africa
  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Vaccines*
  • Bacteriological Techniques / methods*
  • Bronchoalveolar Lavage Fluid / microbiology*
  • Cattle
  • Cattle Diseases / diagnosis*
  • Cattle Diseases / microbiology
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Gene Knockout Techniques
  • Molecular Sequence Data
  • Mycoplasma mycoides / isolation & purification*
  • Oligonucleotide Probes / genetics
  • Pleuropneumonia, Contagious / diagnosis*
  • Pleuropneumonia, Contagious / microbiology
  • Polymerase Chain Reaction / methods*
  • Sequence Analysis, DNA
  • Virulence Factors / deficiency

Substances

  • Bacterial Proteins
  • Bacterial Vaccines
  • DNA, Bacterial
  • Oligonucleotide Probes
  • Virulence Factors

Associated data

  • GENBANK/AM158959