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通过原位聚合酶链反应对组织培养感染细胞中的口蹄疫病毒RNA进行定位

Localization of foot and mouth disease virus RNA in tissue culture infected cells via in situ polymerase chain reaction.

作者信息

Prato Murphy M L, Rodriguez M, Schudel A A, Meyer R F

机构信息

Institute of Virology, CICV-INTA, Morón, Argentina.

出版信息

J Virol Methods. 1995 Aug;54(2-3):173-8. doi: 10.1016/0166-0934(95)00039-w.

Abstract

Foot and mouth disease virus RNA was visualized in infected primary tissue culture cells by in situ PCR incorporating digoxigenin-labeled dUTP. The viral RNA polymerase gene was used as a target for amplification. Infected cells revealed cytoplasmic staining, predominantly perinuclear. The intensity of staining was in proportion to the degree of cytopathology observed and similar to the results obtained using immunoperoxidase staining. The in situ PCR technique for FMDV detection could be applied to formalin-fixed samples and be useful for the study of persistent infections.

摘要

通过掺入地高辛标记的dUTP的原位PCR技术,在感染的原代组织培养细胞中观察到口蹄疫病毒RNA。病毒RNA聚合酶基因用作扩增靶标。受感染的细胞呈现细胞质染色,主要是核周染色。染色强度与观察到的细胞病变程度成正比,且与使用免疫过氧化物酶染色获得的结果相似。用于口蹄疫病毒检测的原位PCR技术可应用于福尔马林固定的样本,对持续性感染的研究很有用。

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