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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (1): 412-414.

所属专题： 生物技术； 畜牧兽医

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

猪圆环病毒的分子生物学检测

谢昆 胡俊杰 全舒舟 高雨蔓   

  • 收稿日期:2010-06-28 修回日期:2010-09-03 出版日期:2011-01-07 发布日期:2011-01-07
  • 基金资助:

    国家科技支撑计划项目

Molecular Biology Detection of Porcine Circovirus

  • Received:2010-06-28 Revised:2010-09-03 Online:2011-01-07 Published:2011-01-07

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摘要：

根据GenBank中发表的猪圆环病毒2型(porcine circovirus type 2, 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573 bp，94°C 30 sec，60°C 30 sec，72°C 30 sec，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25.8 ng；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。

关键词: 红棕象甲, 红棕象甲, 生物学, 防治

Abstract:

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 study 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 is 30 cycles of 94°C 30 sec, 60°C 30 sec, 72°C 30 sec. The template DNA dilution was 100 times the minimum, which can detect the quality of the minimum template DNA was 25.8 ng. The repeatability of PCR program is better. The detection method provides a method and basis of the rapid detection of PCV-2.