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应用逆转录聚合酶链反应(RT-PCR)检测犬外周血单个核细胞中的犬瘟热病毒核衣壳蛋白基因。

Detection of canine distemper virus nucleocapsid protein gene in canine peripheral blood mononuclear cells by RT-PCR.

作者信息

Shin Y, Mori T, Okita M, Gemma T, Kai C, Mikami T

机构信息

Department of Veterinary Microbiology, Faculty of Agriculture, University of Tokyo, Japan.

出版信息

J Vet Med Sci. 1995 Jun;57(3):439-45. doi: 10.1292/jvms.57.439.

Abstract

For a rapid diagnosis of canine distemper virus (CDV) infection, the reverse transcription-PCR (RT-PCR) was carried out to detect CDV nucleoprotein (NP) gene from canine peripheral blood mononuclear cells (PBMCs). Two sets of primers were targeted to two regions of NP gene of CDV Onderstepoort strain. The NP gene fragments were well amplified by the RT-PCR in each of the RNA extracts from Vero cells infected with 6 laboratory strains of CDV including Onderstepoort strain, and from PBMCs of a dog experimentally infected with CDV. The amplified NP gene was detected in 17 of 32 samples from dogs which were clinically suspected for CDV infection at veterinary hospitals. No RT-PCR product was found in 52 samples from healthy dogs including 40 specific pathogen free beagles vaccinated with an attenuated live virus-vaccine for CDV and 12 stray dogs. The RT-PCR provides a fast, sensitive, and supplementary method for the diagnosis of CDV infection in dogs.

摘要

为快速诊断犬瘟热病毒(CDV)感染,采用逆转录聚合酶链反应(RT-PCR)从犬外周血单个核细胞(PBMCs)中检测CDV核蛋白(NP)基因。两组引物靶向CDV Onderstepoort株NP基因的两个区域。在感染了包括Onderstepoort株在内的6种CDV实验室毒株的Vero细胞的RNA提取物以及实验感染CDV的犬的PBMCs中,通过RT-PCR均成功扩增出NP基因片段。在兽医医院临床疑似CDV感染的犬的32份样本中,有17份检测到扩增的NP基因。在52份来自健康犬的样本中未发现RT-PCR产物,其中包括40只接种了CDV减毒活疫苗的无特定病原体比格犬和12只流浪犬。RT-PCR为犬CDV感染的诊断提供了一种快速、灵敏的补充方法。

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