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比较有无RNA提取及病毒分离的聚合酶链反应用于检测犊牛中的牛病毒性腹泻病毒。

A comparison of polymerase chain reaction with and without RNA extraction and virus isolation for detection of bovine viral diarrhea virus in young calves.

作者信息

Deregt D, Carman P S, Clark R M, Burton K M, Olson W O, Gilbert S A

机构信息

Canadian Food Inspection Agency Lethbridge Laboratory, Animal Diseases Research Institute, Lethbridge, Alberta.

出版信息

J Vet Diagn Invest. 2002 Sep;14(5):433-7. doi: 10.1177/104063870201400516.

Abstract

Previously, the authors described a multiplex reverse transcriptase-polymerase chain reaction (PCR) assay for detection and typing of bovine viral diarrhea virus (BVDV) from blood of persistently infected (PI) cattle that could be used with or without RNA extraction. In the present study, the PCR assay was evaluated for its ability to detect BVDV in young calves as a screening tool for detection of persistent infections. Both methods, PCR after RNA extraction (rPCR) and the direct method without RNA extraction (dPCR) were applied and compared with virus isolation (VI) with diagnostic specimens. From 450 whole blood samples from Ontario calves, 47 and 39 samples were positive by rPCR and VI, respectively. From the 47 samples positive by rPCR, 45 (96%) also were positive by dPCR when samples were tested both undiluted and diluted 1:10. In comparison to VI, the relative sensitivities of both PCR assays were 100%. Examination of the results indicates that both PCR assays can be used for screening calves for persistent infection with BVDV.

摘要

此前,作者描述了一种多重逆转录-聚合酶链反应(PCR)检测方法,用于从持续感染(PI)牛的血液中检测牛病毒性腹泻病毒(BVDV)并进行分型,该方法在有无RNA提取的情况下均可使用。在本研究中,对该PCR检测方法作为检测持续性感染的筛查工具在幼龄犊牛中检测BVDV的能力进行了评估。应用了两种方法,即RNA提取后PCR(rPCR)和不进行RNA提取的直接方法(dPCR),并与诊断标本的病毒分离(VI)进行比较。从安大略省犊牛的450份全血样本中,rPCR和VI分别检测出47份和39份阳性样本。在通过rPCR检测为阳性的47份样本中,当样本未经稀释和1:10稀释进行检测时,45份(96%)通过dPCR也呈阳性。与VI相比,两种PCR检测方法的相对灵敏度均为100%。结果检查表明,两种PCR检测方法均可用于筛查犊牛是否感染BVDV持续性感染。

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