复合RT-PCR检测猪乙脑病毒和猪流感病毒方法的建立  

Establishment of Multiplex Reverse Transcription Polymerase Chain Reaction for the Detection of Japanese Encephalitis Virus and Swine Influenza Virus

作　　者：吕晓丽[1] 崔保安[1] 陈红英[1] 魏战勇[1] 王东方[1] 郭小参[1] 

机构地区：[1]河南农业大学牧医工程学院

出　　处：《华南农业大学学报》2008年第4期79-82,共4页Journal of South China Agricultural University

基　　金：“十五”国家食品安全重大攻关专项(2001BA804A30-11);河南省重大科技攻关项目(0223013800)

摘　　要：根据GenBank上已发表的猪乙脑病毒(JEV)的E基因序列和猪流感病毒(SIV)的NP基因序列,设计合成了2对特异引物,分别建立了JEV和SIV的单项RT-PCR诊断方法,通过对扩增条件的筛选,最终成功地建立了JEV和SIV的复合RT-PCR诊断方法,利用一次RT-PCR反应,即可同时扩增JEV的349 bp和SIV 155 bp的特异性片段,而猪瘟病毒(HCV)、猪蓝耳病病毒(PRRSV)、正常细胞(PK-15)核酸扩增结果为阴性,对JEV和SIV的最低检出量分别为100和10 pg的cDNA.该方法适合对JEV和SIV的联合检测和鉴别诊断.Based on the E sequences of JEV and NP sequences of SIV obtained from GenBank^TM, two pairs of primers were designed, single RT-PCR assays for JEV and SIV were established. The multiplex RT-PCR was developed by optimizing amplification conditions. The results showed that fragments of 349 bp for JEV and 155 bp for SIV could be amplified in a multiplex RT-PCR assay. DNA fragments were not amplified from HCV,PRRSV and control culture cells （PK-15）, and this assay could detect the template cDNA of 100 pg for JEV and 10 pg for SIV, and it could also detect JEV, SIV and mixed infection.

关 键 词：复合RT-PCR 检测 猪乙脑病毒 猪流感病毒 

分 类 号：S858.28[农业科学—临床兽医学]