Agriculture and Agri-Food Canada, Food Research and Development Centre, Saint-Hyacinthe, QC, Canada.
J Appl Microbiol. 2010 Jun;108(6):2191-8. doi: 10.1111/j.1365-2672.2009.04624.x. Epub 2009 Nov 14.
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.
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.
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.
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.
本研究旨在开发和优化分子工具，以检测猪和牛体内 Torque teno 病毒（TTV）的存在。开发了一种新型实时聚合酶链反应（PCR），使用 TaqMan 探针来检测两种 TTV 株的基因群。
根据基因组高度保守的非编码区序列分析，设计了寡核苷酸引物和杂交探针。实时 PCR 检测法特异性检测牛和猪 TTV DNA，而不会交叉扩增其他常见病原体。该检测法与常规 PCR 和巢式 PCR 检测法比较，用于检测猪的 1 型和 2 型基因群和牛 TTV 的血浆和粪便样本，该检测法更快、更可靠，降低了假阳性结果的风险。
与常规 PCR 检测法相比，实时 PCR 检测法为猪和牛的两种 TTV 基因群提供了更好的检测结果。
这种新的 TaqMan PCR 检测法将成为检测动物 TTV 株的有用工具，可用于评估动物宿主的病毒载量，最终确定这些病毒在农业食品链中的存在。
