| 猪日本脑炎病毒RT-PCR检测方法的建立 |
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| 引用本文： | 崔奕杰,杨润德,刘晓慧,秦彤,陈丽君,程龙,王琳,苏晓健,孙向华.猪日本脑炎病毒RT-PCR检测方法的建立[J].动物医学进展,2006,27(7):59-62. |
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| 作者姓名： | 崔奕杰 杨润德 刘晓慧 秦彤 陈丽君 程龙 王琳 苏晓健 孙向华 |
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| 作者单位： | 河北农业大学动物科技学院,河北保定,071001 |
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| 摘 要： | 设计了1对引物,利用RT-PCR技术检测乙型脑炎病毒(JEV)。从GenBank中查出收录的31株猪乙型脑炎病毒E基因的已知序列,用DNA star软件对这31株猪乙型脑炎病毒E基因进行同源性分析,以确定扩增的靶序列。以这段靶区域为模板,利用Primer 5软件设计了1对引物,用减毒株SA14-14-2建立了检测乙脑病毒的RT-PCR方法,经敏感性,特异性试验测定,证明该方法敏感,特异;该法可检出样品稀释至256倍的鼠脑毒,相当于0.06个TCID50,对4株河北地区JEV分离株进行检测,结果所设计引物对4株病毒均能扩增出预期的片段。
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| 关 键 词： | 乙型脑炎病毒 反转录-聚合酶链反应 猪 |
| 文章编号： | 1007-5038（2006）07-0059-04 |
| 收稿时间： | 2006-03-27 |
| 修稿时间： | 2006年3月27日 |
Establishment of RT-PCR Detection Method for Japanese encephalitis virus in Swine |
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| CUI Yi-jie,YANG Run-de,LIU Xiao-hui,QIN Tong,CHEN Li-jun,CHENG Long,WANG Lin,SU Xiao-jian,SUN Xiang-hua.Establishment of RT-PCR Detection Method for Japanese encephalitis virus in Swine[J].Progress In Veterinary Medicine,2006,27(7):59-62. |
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| Authors: | CUI Yi-jie YANG Run-de LIU Xiao-hui QIN Tong CHEN Li-jun CHENG Long WANG Lin SU Xiao-jian SUN Xiang-hua |
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| Institution: | College of Animal Science and Veterinary Medicine ,Agriculture University of Hebei , Baoding , Hebei ,071001 , China |
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| Abstract: | We used RT-PCR to detect Japanese encephalitis virus with the designed primers,comparing with the serological method,the former was faster and more sensitive.The homology of the sequences of E genes of 31 strains Japanese encephalitis viruses in GenBank was analyzed by DNA-STAR system.According to the definite target sequences,a pair of primers were designed by Primer 5 system,and RT-PCR was established to detect JEV by attenuated strain SA14-14-2.The results showed that this method was more sensitive and more specific.By the RT-PCR,0.06 TCID_50 JEV could be detected.Four HeBei isolates of JEV were detected by RT-PCR using this pair of primers,and that all the four isolates could be amplified by RT-PCR. |
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| Keywords: | Japanese encephalitis virus RT-PCR Swine |
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