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应用RT-PCR技术快速检测鉴定猪瘟病毒
引用本文:李春阳,单松华,华修国.应用RT-PCR技术快速检测鉴定猪瘟病毒[J].上海交通大学学报(农业科学版),2003,21(1):34-39.
作者姓名:李春阳  单松华  华修国
作者单位:1. 上海师范大学,生命与环境学院,上海,200234
2. 上海出入境检验检疫局,上海,200135
3. 上海交通大学,农业与生物学院,上海,201101
摘    要:根据所有已报道的猪瘟病毒株(HCV)和牛病毒性腹泻病毒株(BVDV)的序列设计了3条引物Pa,Pb,Pc。应用RT-PCR技术,引物对Pa/Pb从猪瘟HCLV株,猪瘟Thiveral株,3株猪瘟野毒株都扩增出了预期185bp的片断,Pa/Pc从BVDVOregonC24V株中也扩增出了240bp的片断,而Pa/Pb与牛睾九原代细胞,PK-15细胞,BVDVOregonC24V株均无反应,Pa/Pc也不与所试验的5株猪瘟毒株反应。采用多重PCR可以从同时含有HCV和BVDV核酸的样品中扩增出各自的特异性条带。针对猪瘟兔化弱毒疫苗株(HCLV)的特异性插入序列。设计了引物Pr,Pra,Prb,可以从HCLV细胞毒或用HCLV人工感染的兔脾组织毒中扩增出预期200bp的片断,而Pr,Pra,Prb与猪瘟Thiveral株和3株猪瘟野毒株均不反应,通过含毒量梯度试验,得出了RT-PCR可检测猪瘟病毒量的下限为200RID或200TCID50。

关 键 词:应用  RT-PCR技术  猪瘟病毒  牛病毒性腹泻病毒
文章编号:1671-9964(2003)01-0034-06
修稿时间:2002年4月17日

Rapid detection of Hog cholera virus by RT- PCR technique
LI Chun-yang,SHAN Song-hu,HUAXiu-guo.Rapid detection of Hog cholera virus by RT- PCR technique[J].Journal of Shanghai Jiaotong University (Agricultural Science),2003,21(1):34-39.
Authors:LI Chun-yang  SHAN Song-hu  HUAXiu-guo
Institution:LI Chun-yang 1,SHAN Song-hua 2,HUAXiu-guo 3
Abstract:Based on the sequence analyses of Hog Cholera Virus(HCV)and Bovine Viral Diarrhea Virus(BVDV),three primers Pa,Pb and Pc were designed and synthesized.The e xpected 185bp fragment was produced by RT-PCR using primer set Pa /Pb from all HCV strains including HCLV,Thiveral and three other wild i solates,whereas using primer set Pa/Pc a 240bp fragment was amplified fr om BVDV Oregon C24V strain.Using multiplex-PCR strategy,these two specific fragments was produce d from the sample mixture containing HCV and BVDV simultaneously. Morever,as for the specific inserte d sequence of HCLV,three other prime rs Pr,Pra and Prb were designed and sy nthesized too.These primers generate a 200bp f ragment by RT-PCR only in the case of H CLV strain.Therefore,they could be used to differentiate HCLV strain from Thiv eral strain and all wild isolates of H CV.The sensitivity of RT-PCR detection for HCV is up to 200RID or 200TCID50
Keywords:HCV  BVDV  RT-PCR  rapid detection
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