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来自南非的牛病毒性腹泻病毒分离株的分子分析。

Molecular analysis of bovine viral diarrhoea virus isolates from South Africa.

作者信息

Kabongo N, Baule C, Van Vuuren M

机构信息

Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Private Bag X04, Onderstepoort, 0110 South Africa.

出版信息

Onderstepoort J Vet Res. 2003 Dec;70(4):273-9. doi: 10.4102/ojvr.v70i4.292.

Abstract

The presence of bovine viral diarrhoea virus in South Africa has been confirmed by several serological surveys. However, little is known about its biological properties. Twenty five isolates obtained by isolation in tissue culture and detected by means of the antigen capture ELISA from clinically sick cattle and from foetal calf serum in South Africa were characterized on the basis of analysis of the 5' non-translated (NTR) region of the genome. A reverse-transcription polymerase chain reaction (RT-PCR) was used to amplify specific sequences from the 5'NTR of the genome. The oligonucleotide primers corresponding to positions 105-125 and 399-378, respectively, in the sequence of BVDV strain NADL were used to generate the PCR products. Both strands were sequenced directly with these primers and fluorescence-labelled dideoxynucleotides in an automated nucleic acid sequencer. Reference strains of pestiviruses [(BVDV type I, BVDV type II, border disease virus (BDV) and hog cholera virus (HCV)] and isolates from a previous investigation on BVDV in southern Africa were included for comparative purposes. All the BVDV strains obtained during this study belong to subgroups of BVDV genotype I. No association could be demonstrated between the geographic origin of the isolates. A number of isolates formed another branch separate from the existing branches Ia, Ib and Ic. These findings suggest that extensive genetic diversity can be found within BVDV type I isolates from southern Africa. Isolates that group with the classical BVDV type I strains, particularly of American origin, coexist with variants that appear to represent a local genetic pool and or variants evolving from the classical strains.

摘要

多项血清学调查已证实南非存在牛病毒性腹泻病毒。然而,对其生物学特性却知之甚少。通过组织培养分离并利用抗原捕获酶联免疫吸附测定法(ELISA)从南非临床患病牛和胎牛血清中检测到的25株病毒,基于对基因组5'非翻译(NTR)区域的分析进行了特征鉴定。采用逆转录聚合酶链反应(RT-PCR)从基因组的5'NTR扩增特定序列。分别对应于牛病毒性腹泻病毒(BVDV)NADL株序列中第105 - 125位和399 - 378位的寡核苷酸引物用于生成PCR产物。两条链均使用这些引物和荧光标记的双脱氧核苷酸在自动核酸测序仪中直接测序。为了进行比较,纳入了瘟病毒的参考毒株[(I型BVDV、II型BVDV、边界病病毒(BDV)和猪霍乱病毒(HCV)]以及先前在南部非洲对BVDV进行调查时获得的分离株。本研究期间获得的所有BVDV毒株均属于I型BVDV基因型的亚组。分离株的地理来源之间未显示出关联。一些分离株形成了一个与现有Ia、Ib和Ic分支分开的另一个分支。这些发现表明,在来自南部非洲的I型BVDV分离株中可发现广泛的遗传多样性。与经典I型BVDV毒株(特别是美国来源的)归为一组的分离株,与似乎代表本地基因库和/或从经典毒株演变而来的变体共存。

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