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建立一种实时 TaqMan PCR 检测方法用于检测猪和牛的扭曲线粒体病毒。

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

机构信息

Agriculture and Agri-Food Canada, Food Research and Development Centre, Saint-Hyacinthe, QC, Canada.

出版信息

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.

摘要

目的

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。

方法和结果

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。

结论

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。

研究的意义和影响

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。

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