Development of a real-time TaqMan PCR assay for the detection of porcine and bovine Torque teno virus

J Appl Microbiol. 2010 Jun;108(6):2191-8. doi: 10.1111/j.1365-2672.2009.04624.x. Epub 2009 Nov 14.

Abstract

Aims: The goal of this study was to develop and to optimize molecular tools to detect the presence of Torque teno virus (TTV) in swine and cattle. A novel real-time polymerase chain reaction (PCR) using a TaqMan probe was developed to detect both genogroups of TTV strains.

Methods and results: Oligonucleotide primers and hybridization probes were designed based on sequence analysis of the noncoding region, a highly conserved part of the genome. The real-time PCR assay specifically detected bovine and porcine TTV DNA without cross-amplification of other common pathogens. The assay was compared with conventional PCR and nested-PCR assays for the detection of porcine genogroups 1 and 2 and bovine TTV on plasma and faecal samples, and the assay was found faster, more reliable and reduced the risk of false positive results.

Conclusions: The real-time PCR assay provided better detection results for the two TTV genogroups in both swine and cattle compared to the conventional PCR assays.

Significance and impact of the study: This new TaqMan PCR assay will be a useful tool for the detection of animal TTV strains, to evaluate the viral load from animal host and finally to identify the presence of these viruses in the agri-food continuum.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cattle / virology
  • Cattle Diseases / virology*
  • DNA Primers
  • DNA Virus Infections / veterinary*
  • DNA Virus Infections / virology
  • DNA, Viral / isolation & purification
  • Feces / virology
  • Molecular Sequence Data
  • Oligonucleotide Probes
  • Phylogeny
  • Plasma / virology
  • Polymerase Chain Reaction / methods*
  • Polymerase Chain Reaction / veterinary
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Species Specificity
  • Swine / virology
  • Swine Diseases / virology*
  • Torque teno virus / genetics
  • Torque teno virus / isolation & purification*

Substances

  • DNA Primers
  • DNA, Viral
  • Oligonucleotide Probes