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一种用于检测和鉴定水疱性口炎病毒核酸的半巢式聚合酶链反应检测法。

A hemi-nested PCR assay for the detection and identification of vesicular stomatitis virus nucleic acid.

作者信息

Höfner M C, Carpenter W C, Ferris N P, Kitching R P, Ariza Botero F

机构信息

AFRC Institute for Animal Health, Pirbright Laboratory, Woking, Surrey, UK.

出版信息

J Virol Methods. 1994 Dec;50(1-3):11-20. doi: 10.1016/0166-0934(94)90159-7.

Abstract

This paper is the first to describe the development of a hemi-nested PCR assay for the detection of vesicular stomatitis virus (VSV) nucleic acid. This assay was developed as it combines high sensitivity for virus genome detection with the identification of the external amplification product in the reamplification step, thus confirming the specificity of the reaction. The assay did not depend on the presence of infectious virus in samples, as demonstrated by its detection of VSV in blood samples which were non-infectious in tissue culture. One further advantage was that the VSV-New Jersey and VSV-Indiana serotypes could be differentiated through the selective use of the appropriate hemi-nested primer. This assay is ideal for the study of VSV pathogenesis and persistence.

摘要

本文首次描述了一种用于检测水疱性口炎病毒(VSV)核酸的半巢式PCR检测方法的开发。开发该检测方法是因为它将病毒基因组检测的高灵敏度与再扩增步骤中外部扩增产物的鉴定相结合,从而确认了反应的特异性。该检测方法不依赖于样本中感染性病毒的存在,这在其对组织培养中无感染性的血液样本中的VSV检测中得到了证明。另一个优点是,通过选择性使用合适的半巢式引物,可以区分VSV-新泽西型和VSV-印第安纳型血清型。该检测方法对于研究VSV的发病机制和持续性非常理想。

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