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使用核酸杂交技术检测感染牛病毒性腹泻病毒的牛。

Detection of cattle infected with bovine viral diarrhea virus using nucleic acid hybridization.

作者信息

Roberts K L, Collins J K, Carman J, Blair C D

机构信息

Department of Microbiology, Colorado State University, Fort Collins 80523.

出版信息

J Vet Diagn Invest. 1991 Jan;3(1):10-5. doi: 10.1177/104063879100300103.

Abstract

A ribonucleic acid (RNA) hybridization assay to identify cattle infected by bovine viral diarrhea virus (BVDV) is described. The RNA probe was derived from the coding region at the 3' end of the genome of the NADL strain of BVDV. Total RNA from infected cell cultures or peripheral blood leukocytes from suspect animals was extracted and applied to nylon membranes with a slot blot apparatus. Peripheral blood leukocytes were tested concurrently for BVDV by virus isolation. The results of hybridization and virus isolation were in agreement for 92% of the cases. When compared with virus isolation, hybridization had a sensitivity of detection of 59.5% and a specificity of 95%. Cross-reactivity to RNA extracts of border disease virus-infected cells was noted. No cross-reactivity was detected to other common bovine viruses (bovine herpesvirus-1, bovine respiratory syncytial virus, parainfluenza-3 virus, and bluetongue virus), to viruses classified in related families (equine arteritis virus and Venezuelan equine encephalitis virus), or to viruses having similar genomic organization (dengue virus type 2 and Japanese encephalitis virus).

摘要

本文描述了一种用于鉴定感染牛病毒性腹泻病毒(BVDV)的牛的核糖核酸(RNA)杂交试验。RNA探针源自BVDV NADL株基因组3'端的编码区。从感染的细胞培养物或疑似动物的外周血白细胞中提取总RNA,并使用狭缝印迹装置将其应用于尼龙膜。同时通过病毒分离对外周血白细胞进行BVDV检测。在92%的病例中,杂交结果与病毒分离结果一致。与病毒分离相比,杂交检测的灵敏度为59.5%,特异性为95%。注意到与边境病病毒感染细胞的RNA提取物有交叉反应。未检测到与其他常见牛病毒(牛疱疹病毒1型、牛呼吸道合胞病毒、副流感3型病毒和蓝舌病毒)、相关科分类的病毒(马动脉炎病毒和委内瑞拉马脑炎病毒)或具有相似基因组组织的病毒(2型登革病毒和日本脑炎病毒)的交叉反应。

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