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一种新型的滚环扩增检测方法,用于检测猪和人类圆环病毒科的成员。

A novel rolling circle amplification assay to detect members of the family Anelloviridae in pigs and humans.

机构信息

Centre de Recerca en Sanitat Animal, UAB-IRTA, Campus de Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain.

出版信息

Virus Res. 2011 Sep;160(1-2):424-7. doi: 10.1016/j.virusres.2011.06.025. Epub 2011 Jul 6.

Abstract

The study describes a novel rolling circle amplification (RCA) assay to detect members of the family Anelloviridae from swine and human serum samples. The RCA was carried out using short anellovirus (AV)-specific primers based on a highly conserved region among available AV full-length genomes. Then, RCA products were used as templates to amplify full-length genomes with an AV-specific PCR. Amplification products were separated by agarose gel electrophoresis and full-length genomes were isolated based on the known size. With this novel AV-RCA/PCR approach it was possible to detect Torque teno sus virus 1 (TTSuV1) and Torque teno sus virus 2 (TTSuV2) in swine and many species of Torque teno virus (TTV) in human sera, which were previously tested negative by conventional PCRs.

摘要

本研究描述了一种新颖的滚环扩增(RCA)检测方法,用于检测来自猪和人血清样本的圆环病毒科成员。该 RCA 是使用基于现有圆环病毒全长基因组之间高度保守区域的短圆环病毒(AV)特异性引物进行的。然后,将 RCA 产物用作模板,使用 AV 特异性 PCR 扩增全长基因组。通过琼脂糖凝胶电泳分离扩增产物,并根据已知大小分离全长基因组。通过这种新颖的 AV-RCA/PCR 方法,可以检测到猪中的 Torque teno sus 病毒 1(TTSuV1)和 Torque teno sus 病毒 2(TTSuV2),以及人血清中的多种 Torque teno 病毒(TTV),而这些病毒之前用常规 PCR 检测为阴性。

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