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根据中国的系统发育关系,用反转录-聚合酶链反应-限制性片段长度多态性区分来自不同疫苗株的毛皮动物犬瘟热病毒分离株。

Differentiation of canine distemper virus isolates in fur animals from various vaccine strains by reverse transcription-polymerase chain reaction-restriction fragment length polymorphism according to phylogenetic relations in china.

机构信息

Jilin Zhongte Biotechnological Co, Ltd, Jilin 132109, People's Republic of China.

出版信息

Virol J. 2011 Feb 27;8:85. doi: 10.1186/1743-422X-8-85.

Abstract

In order to effectively identify the vaccine and field strains of Canine distemper virus (CDV), a new differential diagnostic test has been developed based on reverse transcription-polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP). We selected an 829 bp fragment of the nucleoprotein (N) gene of CDV. By RFLP analysis using BamHI, field isolates were distinguishable from the vaccine strains. Two fragments were obtained from the vaccine strains by RT-PCR-RFLP analysis while three were observed in the field strains. An 829 nucleotide region of the CDV N gene was analyzed in 19 CDV field strains isolated from minks, raccoon dogs and foxes in China between 2005 and 2007. The results suggest this method is precise, accurate and efficient. It was also determined that three different genotypes exist in CDV field strains in fur animal herds of the north of China, most of which belong to Asian type. Mutated field strains, JSY06-R1, JSY06-R2 and JDH07-F1 also exist in Northern China, but are most closely related to the standard virulent strain A75/17, designated in Arctic and America-2 genetype in the present study, respectively.

摘要

为了有效鉴别犬瘟热病毒(CDV)的疫苗株和野毒株,我们基于反转录-聚合酶链反应(RT-PCR)和限制性片段长度多态性(RFLP)开发了一种新的鉴别诊断检测方法。我们选择了 CDV 的核蛋白(N)基因的 829 个核苷酸片段。通过 BamHI 的 RFLP 分析,野毒株与疫苗株可区分开来。通过 RT-PCR-RFLP 分析,从疫苗株中获得了两个片段,而在野毒株中观察到了三个片段。对 2005 年至 2007 年间在中国水貂、貉和狐狸中分离的 19 株 CDV 野毒株的 CDV N 基因的 829 个核苷酸区域进行了分析。结果表明,该方法精确、准确、高效。还确定了在中国北方毛皮动物养殖场的 CDV 野毒株中存在三种不同的基因型,其中大多数属于亚洲型。在中国北方也存在突变的野毒株 JSY06-R1、JSY06-R2 和 JDH07-F1,但与本研究中指定的北极和美洲-2 基因型的标准强毒 A75/17 最为密切相关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5f2/3056815/d35eef16b8d0/1743-422X-8-85-1.jpg

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