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羊痘病毒多重PCR检测方法的建立
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韩若婵,张强*，吴国华，颜新敏，李健，朱海霞，朱彩珠，
谢芳，鞠厚斌，施程洪，才学鹏*
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（中国农业科学院 兰州兽医研究所 家畜疫病病原生物学国家重点实验室 农业部畜禽病毒学重点
开放实验室,甘肃 兰州730046）
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摘要：
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根据羊痘病毒(CaPV)末端反向重复序列中的一段序列和P32基因序列，设计合成了2对引物，通过确定最佳的PCR反应条件，建立了检测羊痘病毒核酸的多重PCR方法。结果显示，用这2对引物均能扩增出羊痘病毒相应的特异性片段，而用此PCR方法检测口蹄疫病毒和羊口疮病毒，结果均为阴性。与中和试验比较，建立的PCR方法具有更好的敏感性。表明，该PCR方法可对组织病料中的CaPV进行快速检测。 |
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关键词：
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羊痘病毒；多重PCR；特异性；敏感性 |
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中图分类号：S 852.659.1：Q 503 文献标识码：
A 文章编号：1673-4696(2008)03-0206-03 |
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Development of a multiplex PCR assay for detection of capripoxvirus |
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HAN Ruo-chan,ZHANG Qiang,WU Guo-hua,YAN Xin-min,LI Jian,ZHU Hai-xia,ZHU Cai-zhu,
XIE Fang,JU Hou-bin,SHI Cheng-hong,CAI Xue-peng |
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(Key Laboratory of Animal Virology of the Ministry of Agriculture/State Key Laboratory of Veterinary Etiological
Biology/ Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China) |
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Abstract: Two pairs of specific primers were designed and synthesized according to the inverted terminal repeat sequence and p32 gene sequence of the genome of capripoxvirus(CaPV).After optimization of PCR conditions,a multiplex PCR assay for detecting the genomic DNA of CaPV was developed.The results revealed that the expected fragment could be amplified from the extracted CaPV DNA by the multiplex PCR assay developed.No PCR products were amplified from samples containing orf virus or foot-and-mouth disease virus using the assay.Compared with the neutralization tests,the multiplex PCR was more sensitive,and would be applied for rapid diagnosis of CaPV samples in biopsy and tissue cultures.
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Key words: capripoxvirus；multiplex PCR；specificity；sensitivity |
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