环介导恒温扩增法检测卡氏肺孢子菌方法的建立

中国寄生虫学与寄生虫病杂志 ›› 2010, Vol. 28 ›› Issue (4): 18-307.

环介导恒温扩增法检测卡氏肺孢子菌方法的建立

张辉，刘雪晴，何荣志，贾天军，张进顺*

河北北方学院病原生物学及免疫学研究所，张家口 075000

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-08-30 发布日期:2010-08-30

Establishment of Loop-mediated Isothermal Amplification （LAMP） for Detecting Pneumocystis carinii

ZHANG Hui，LIU Xue-qing，HE Rong-zhi，JIA Tian-jun，ZHANG Jin-shun*

Institute of Pathogenic Biology and Immunology，Hebei North University，Zhangjiakou 075000，China

Received:1900-01-01 Revised:1900-01-01 Online:2010-08-30 Published:2010-08-30

摘要/Abstract

摘要： 通过建立肺孢子菌肺炎动物模型获取感染卡氏肺孢子菌（Pneumocystis carinii）的大鼠肺组织，用环介导恒温扩增法（LAMP）扩增卡氏肺孢子菌核内核糖体小亚基18s rDNA，扩增产物用限制性内切酶ApalⅠ酶切，观察酶切片段大小。卡氏肺孢子菌核内核糖体小亚基18s rDNA克隆至载体pGEX6p2，筛选阳性克隆。本研究初步建立了检测卡氏肺孢子菌的环介导恒温扩增法。

关键词: 卡氏肺孢子菌, 18s rDNA, 环介导恒温扩增

Abstract: Animal model of Pneumocystis carinii pneumonia （PCP） was established for acquiring lung tissue infected with P. carinii. After DNA from rat lungs was extracted， nuclear ribosome small subumit 18s rDNA of P. carinii was amplified by loop-mediated isothermal amplification method at 63 ℃ for 60 min. The product was digested by restriction enzyme ApalⅠ. The results showed that 18s ribosome DNA （rDNA） of P. carinii was cloned into vector pGEX6p2， and the positive clones screened. Therefore, the loop-mediated isothermal amplification has been established for detecting P. carinii.

Key words: Pneumocystis carinii, 18s rDNA, Loop-mediated isothermal amplification

张辉;刘雪晴;何荣志;贾天军;张进顺. 环介导恒温扩增法检测卡氏肺孢子菌方法的建立[J]. 中国寄生虫学与寄生虫病杂志, 2010, 28(4): 18-307.

ZHANGHui;LIUXue-qing;HERong-zhi;JIATian-jun;ZHANGJin-shun*. Establishment of Loop-mediated Isothermal Amplification （LAMP） for Detecting Pneumocystis carinii[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2010, 28(4): 18-307.

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