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高致病性猪繁殖与呼吸综合征病毒RT—PCR鉴别诊断方法的建立

ISSN号：1008-0589
期刊名称：《中国预防兽医学报》
时间：0
分类：S852.659.6[农业科学—基础兽医学;农业科学—兽医学;农业科学—畜牧兽医] S854.44[农业科学—临床兽医学;农业科学—兽医学;农业科学—畜牧兽医]
作者机构：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室／猪传染病研究室,黑龙江哈尔滨150001
相关基金：国家“973”计划（2005CB523200）;十一五国家科技支撑计划（2006BAD06A03/01/04）;国家自然科学基金资助项目（30470072）

作者：郝晓芳[1], 周艳君[1], 田志军[1], 韦天超[1], 安同庆[1], 彭金美[1], 华荣虹[1], 童光志[1]

关键词：高致病性猪繁殖与呼吸综合征病毒, 基因缺失, RT-PCR, 鉴别诊断, highly pathogenic PRRSV, gene deletion, RT-PCR, differential diagnosis

中文摘要：

2006年5月以来，我国部分猪场暴发了一种以高热、高发病率和高死亡率为特征的传染性疾病，经病原分离及分子流行病学分析证明是由一种带有nsp2部分缺失的、对猪呈高致病性的猪繁殖与呼吸综合征病毒（Porcine reproductive and respiratory syndrome virus，PRRsv）引起的。本实验参考GenBank发表的以及本实验室分离鉴定的PRRSV的nsp2基因序列，在nsp2缺失区的两端的保守区设计并合成了一对引物，建立了一种PRRSV的RT-PCR检测方法。该方法扩增高致病性PRRSV基因组时可获得230bp的片段，扩增经典型PRRSV时则获得320bp的片段，根据RT-PCR产物大小可将二者区分开来。通过大量临床病料的检测，并配合PCR产物测序验证，结果表明该方法简便、快速、特异，可以鉴别高致病性PRRSV，为进一步的PRRS流行病学研究提供了重要的技术手段。

英文摘要：

Since May of 2006, serious outbreaks of a highly contagious disease marked by high fever and high morbidity and mortality have occurred in China. Etiology studies confirmed that the disease was caused by a highly pathogenic porcine reproductive and respiratory syndrome virus （PRRSV） characterized by two non-continued deletions in the nsp2 region of viral genome. In the study, a RT-PCR was developed using a pair of primers designed according to the nsp2 gene sequence of PRRSVs deposited in GenBank and the highly pathogenic PRRSVs isolated in our laboratory. This method was shown to specifically amplify a 230 bp fragment from the highly pathogenic PRRSVs or a 320 bp fragment from the classical form of PRRSVs. Test on a large number of clinical samples showed that this method was highly specific and can be rapidly performed in a standard PCR setting. Therefore the RT-PCR could be used as an effective tool for differentiating diagnosis of the highly pathogenic PRRSV in epidemiological investigations.

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