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


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.


CONCLUSIONS
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.
Brassard
J
J
Agriculture and Agri-Food Canada, Food Research and Development Centre, Saint-Hyacinthe, QC, Canada. julie.brassard@agr.gc.ca
Gagné
M-J
MJ
Houde
A
A
Poitras
E
E
Ward
P
P
D000818:Animals; D001483:Base Sequence; D002417:Cattle / Q000821:virology; D002418:Cattle Diseases / Q000821:virology*; D017931:DNA Primers; D004266:DNA Virus Infections / Q000662:veterinary* / Q000821:virology; D004279:DNA, Viral / Q000302:isolation & purification; D005243:Feces / Q000821:virology; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010802:Phylogeny; D010949:Plasma / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000662:veterinary; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D013552:Swine / Q000821:virology; D013553:Swine Diseases / Q000821:virology*; D022783:Torque teno virus / Q000235:genetics / Q000302:isolation & purification*
D017931:DNA Primers; D004279:DNA, Viral; D015345:Oligonucleotide Probes