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

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

Author information

1
Agriculture and Agri-Food Canada, Food Research and Development Centre, Saint-Hyacinthe, QC, Canada. julie.brassard@agr.gc.ca

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.

[Indexed for MEDLINE]
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