| 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 |
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| 引用本文： | 郝晓芳,周艳君,田志军,韦天超,安同庆,彭金美,华荣虹,童光志.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[J].中国预防兽医学报,2007,29(9):704-709. |
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| 作者姓名： | 郝晓芳 周艳君 田志军 韦天超 安同庆 彭金美 华荣虹 童光志 |
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| 作者单位： | 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 |
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| 基金项目： | 国家重点基础研究发展计划(973计划);国家科技支撑计划;国家自然科学基金 |
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| 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。
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| 关 键 词： | 高致病性猪繁殖与呼吸综合征病毒 基因缺失 鉴别诊断 |
| 文章编号： | 1008-0589（2007）09-0704-06 |
| 修稿时间： | 2007-05-24 |
Development of a RT-PCR method for differentiation of the highly pathogenic PRRSVs and the classical PRRSVs |
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| HAO Xiao-fang,ZHOU Yan-jun,TIAN Zhi-jun,WEI Tian-chao,AN Tong-qing,PENG Jin-mei,HUA Rong-hong,TONG Guang-zhi.Development of a RT-PCR method for differentiation of the highly pathogenic PRRSVs and the classical PRRSVs[J].Chinese Journal of Preventive Veterinary Medicine,2007,29(9):704-709. |
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| Authors: | HAO Xiao-fang ZHOU Yan-jun TIAN Zhi-jun WEI Tian-chao AN Tong-qing PENG Jin-mei HUA Rong-hong TONG Guang-zhi |
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| Institution: | Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China |
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| Abstract: | 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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| Keywords: | RT-PCR |
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