›› 2011, Vol. 38 ›› Issue (7): 84-86.

• 生物技术 • 上一篇    下一篇

以鸡毒支原体pvpA基因序列建立的套式PCR检测方法

毛晓娜, 王嵩林, 徐丽丽, 周莉, 姚琦, 张映   

  1. 山西农业大学动物科技学院,山西太谷 030801
  • 收稿日期:2010-12-27 修回日期:1900-01-01 出版日期:2011-07-20 发布日期:2011-07-20
  • 通讯作者: 张映

Establishment of an Nested PCR for the Detection of MG by pvpA Gene Sequence

MAO Xiao-na, WANG Song-lin, XU Li-li, ZHOU Li, YAO Qi, ZHANG Ying   

  1. College of Animal Science and Veterinary Medicine,Shanxi Agricultural University,Taigu 030801,China
  • Received:2010-12-27 Revised:1900-01-01 Online:2011-07-20 Published:2011-07-20

摘要: 本试验以GenBank中登录的鸡毒支原体(Mycoplasma gallisepticum,MG)的特异性黏附蛋白pvpA基因序列为目标,用两对引物对鸡毒支原体DNA进行套式PCR扩增,建立套式PCR扩增体系,并进行了套式PCR的敏感性和特异性试验。结果显示,应用该PCR方法对MG DNA的检出限为0.18 pg/μL;以鸡常见细菌、病毒DNA为模板进行PCR扩增,均未扩增出条带,说明该方法特异性强,适用于临床对MG早期感染的检测。

关键词: 鸡毒支原体; 套式PCR; 黏附蛋白PvpA; 检测

Abstract: In this experiment,the system and condition of nested PCR to detect MG were established with two pairs of primers which had been designed by the predecessors according to the adherin protein pvpA gene sequence of MG published in GenBank.Theremore, the sensitivity and specificity of this method was analysed.The results showed that this method could detect more than 0.18 pg/μL MG DNA.However, based on these two pairs of internal and external primers, the PCR did not amplify the specific fragment with the template DNA from common bacterium and virus of chicken. The results showed that this PCR amplification system was specific and sensitive. It can be used to detect early stages of MG infection in clinical diagnosis.

Key words: MG; nested PCR; adherin protein-PvpA; detection

中图分类号: