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使用合成寡核苷酸检测牛病毒性腹泻病毒

Detection of BVD viruses using synthetic oligonucleotides.

作者信息

Lewis T L, Ridpath J F, Bolin S R, Berry E S

机构信息

Department of Veterinary Science, North Dakota State University, Fargo.

出版信息

Arch Virol. 1991;117(3-4):269-78. doi: 10.1007/BF01310770.

Abstract

This study examined synthetic oligonucleotide probes as potential diagnostic tools for bovine viral diarrhea virus (BVDV). Six 20-base sequences from across the genome were selected by homology analysis of the published genomic sequences of the NADL and Osloss isolates of BVDV. RNA was extracted from 22 BVDV isolates propagated in bovine turbinate (BT) cells, blotted, and probed with 32P end-labeled oligonucleotides. The stringency conditions used were such that more than a single base mismatch would result in no hybridization. The probe originating nearest the 5' end of the viral RNA, ND001, detected 86% of the viral isolates while the other probes detected from 19% to 57%. Both cytopathic and noncytopathic isolates were detected by these synthetic probes. A cocktail of these probes were used to specifically detect BVDV RNA extracted directly from tissues of cattle either persistently or acutely infected.

摘要

本研究检测了合成寡核苷酸探针作为牛病毒性腹泻病毒(BVDV)潜在诊断工具的情况。通过对已发表的BVDV NADL和Osloss分离株基因组序列进行同源性分析,从整个基因组中选择了6个20碱基序列。从在牛鼻甲(BT)细胞中增殖的22株BVDV分离株中提取RNA,进行印迹,并用32P末端标记的寡核苷酸进行探测。所使用的严谨条件是,超过一个碱基错配将导致无杂交。源自病毒RNA 5'端最近处的探针ND001检测到86%的病毒分离株,而其他探针检测到的比例为19%至57%。这些合成探针检测到了致细胞病变和非致细胞病变的分离株。使用这些探针的混合物特异性检测直接从持续或急性感染牛的组织中提取的BVDV RNA。

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