Glycoprotein C gene based molecular subtyping of a bovine herpesvirus -1 isolate from uttar pradesh, India.

Indian Journal of Virology Pub Date : 2012-12-01 Epub Date: 2012-10-12 DOI:10.1007/s13337-012-0116-x
Chintu Ravishankar, S Nandi, V Chander, T K Mohapatra
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Abstract

Bovine herpesvirus -1 (BHV-1) is the etiological agent of many clinical syndromes in cattle which causes huge economic losses to the animal husbandry sector annually. Since the first report of its presence in India in 1976, the disease is considered to be endemic in the country. In the present study, a case of keratoconjunctivitis in a cow was investigated to find out the underlying cause of the condition. The clinical material (ocular swab) was tested by BHV-1 glycoprotein D gene specific PCR using in house designed primers and found to be positive by the presence of a 212 bp DNA product in agarose gel electrophoresis. The virus was isolated in MDBK cell line in the third passage and the serum from the animal, was positive for antibodies against BHV-1 by ELISA. A 575 bp segment of the glycoprotein C gene of the isolate was amplified by PCR, cloned and sequenced. On phylogenetic analysis, it was seen that the sequence matched with published BHV-1.1 sequences from USA and Uruguay whereas it was divergent from Brazilian BHV-1.1 isolates. This study highlights the isolation, rapid and sensitive detection of BHV-1 virus from clinical cases and its subtyping by nucleotide sequencing and subsequent phylogenetic analysis which gives invaluable information about the molecular epidemiology of BHV-1 subtypes prevalent in the country.

基于糖蛋白 C 基因的印度乌塔尔邦牛疱疹病毒-1 分离物分子亚型鉴定。
牛疱疹病毒-1(BHV-1)是许多牛临床综合症的病原体,每年给畜牧业造成巨大的经济损失。自 1976 年首次报告该病在印度出现以来,该病就被认为是印度的地方病。本研究调查了一例奶牛角结膜炎病例,以找出该病的根本原因。临床材料(眼拭子)通过使用内部设计的引物进行 BHV-1 糖蛋白 D 基因特异性 PCR 检测,在琼脂糖凝胶电泳中发现 212 bp DNA 产物呈阳性。在 MDBK 细胞系中分离出第三代病毒,用 ELISA 检测动物血清中的 BHV-1 抗体呈阳性。通过 PCR 扩增了分离株糖蛋白 C 基因的 575 bp 片段,并对其进行了克隆和测序。系统进化分析显示,该序列与美国和乌拉圭已发表的 BHV-1.1 序列一致,而与巴西的 BHV-1.1 分离物存在差异。这项研究强调了从临床病例中分离、快速和灵敏地检测 BHV-1 病毒,并通过核苷酸测序和随后的系统发育分析对其进行亚型鉴定,从而为巴西流行的 BHV-1 亚型的分子流行病学提供了宝贵信息。
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来源期刊
Indian Journal of Virology
Indian Journal of Virology 医学-病毒学
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6-12 weeks
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