Development of a real-time TaqMan PCR assay for the detection of porcine and bovine Torque teno virus
| Autor: | Pierre Ward, Elyse Poitras, Alain Houde, Julie Brassard, Marie-Josée Gagné |
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| Rok vydání: | 2009 |
| Předmět: |
Torque teno virus
Sequence analysis Swine Pcr assay Molecular Sequence Data Cattle Diseases Biology Applied Microbiology and Biotechnology Genome Polymerase Chain Reaction law.invention Feces Plasma Species Specificity law TaqMan Animals Polymerase chain reaction Phylogeny DNA Primers Swine Diseases Base Sequence General Medicine Sequence Analysis DNA Virology Molecular biology DNA Virus Infections Oligonucleotide primers DNA Viral Cattle Oligonucleotide Probes Viral load Sequence Alignment Biotechnology |
| Zdroj: | Journal of applied microbiology. 108(6) |
| ISSN: | 1365-2672 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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