猪圆环病毒的分子生物学检测  

Molecular Biology Detection of Porcine Circovirus

作　　者：谢昆[1] 胡俊杰[1,2] 全舒舟[1] 高雨蔓[1] 

机构地区：[1]红河学院生命科学与技术学院,云南蒙自661100 [2]云南大学生命科学学院,昆明650091

出　　处：《中国农学通报》2011年第1期412-414,共3页Chinese Agricultural Science Bulletin

基　　金：红河学院重点学科建设项目"生物化学与分子生物学"(071010);红河学院博硕科研启动项目"鸡IL-2基因的克隆和表达"(XS205026)

摘　　要：根据GenBank中发表的猪圆环病毒2型(porcine circovirus type2,PCV-2)基因组序列,利用Primier5设计了一对特异性引物,分别采用CTAB法和DNA试剂盒提取猪脾脏总DNA,用PCR方法扩增猪PCV-2基因保守区序列,并对PCR扩增程序进行优化,对PCR检测方法的敏感性、重复性做了研究。结果显示扩增的目的片段约为573bp,94℃30sec,60℃30sec,72℃30sec,30个循环扩增效果最好;模板DNA最低稀释倍数为100倍,即能检测到最低模板DNA的质量为25.8ng;同一组织不同部位PCR检测的重复性较好;该检测方法为PCV-2的快速检测提供了方法和依据。According to the genomic sequence of PCV-2（porcine circovirus type 2） published in GenBank, which designed a pair of specific primers by using Primier5. The total DNA of Pig spleen extracted by CTAB and DNA Kit. And the PCV-2 gene conserved sequence of Pig is amplificated by PCR. Then we studied the optimization of the PCR process, and proceduring on the PCR sensitivity and repeatability. The result showed that amplified fragment was about 573 bp. The best PCR program was 30 cycles of 94℃ 30 sec, 60℃ 30 sec, 72℃ 30 sec. The template DNA dilution was 100 times the minimum, which could detect the quality of the minimum template DNA was 25.8 ng. The repeatability of PCR program was better. The detection method provides a method and basis of the rapid detection of PCV-2.

关 键 词：猪圆环病毒2型 聚合酶链式反应 检测 

分 类 号：S852.65[农业科学—基础兽医学]