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[建立用于检测卡氏肺孢子虫的环介导等温扩增技术(LAMP)]

[Establishment of loop-mediated isothermal amplification (LAMP) for detecting Pneumocystis carinii].

作者信息

Zhang Hui, Liu Xue-qing, He Rong-zhi, Jia Tian-jun, Zhang Jin-shun

机构信息

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

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2010 Aug;28(4):305-7.

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 subunit 18s rDNA of P. carinii was amplified by loop-mediated isothermal amplification method at 63 degrees C for 60 min. The product was digested by restriction enzyme Apal I. 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.

摘要

为获取卡氏肺孢子虫感染的肺组织,建立了卡氏肺孢子虫肺炎(PCP)动物模型。从大鼠肺中提取DNA后,采用环介导等温扩增法在63℃下对卡氏肺孢子虫的核糖体小亚基18s rDNA进行扩增60分钟。产物用限制性内切酶Apal I消化。结果表明,卡氏肺孢子虫的18s核糖体DNA(rDNA)被克隆到载体pGEX6p2中,并筛选出阳性克隆。因此,已建立环介导等温扩增法用于检测卡氏肺孢子虫。

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