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建立细粒棘球绦虫[已更正]共检测的环介导等温扩增方法(LAMP)。

The development of a loop-mediated isothermal amplification method (LAMP) for Echinococcus granulosus [corrected] coprodetection.

机构信息

Department of Molecular Genetics, Hebrew University-Hadassah Medical School, Jerusalem, Israel.

出版信息

Am J Trop Med Hyg. 2012 Nov;87(5):883-7. doi: 10.4269/ajtmh.2012.12-0184. Epub 2012 Sep 17.

Abstract

We have previously developed a polymerase chain reaction (PCR) assay for detection of Echinococcus granulosus infection, which proved very sensitive and specific for identification of infected dogs. We have now developed a loop-mediated isothermal amplification (LAMP) assay, which amplifies the same genomic repeated sequences of E. granulosus for coprodetection. This assay enabled detection of a single egg in fecal samples and showed high species specificity for E. granulosus with no cross-amplification of DNA from closely related helminths, including Echinococcus multilocularis. Because the method does not require thermocycling for DNA amplification, or electrophoresis for amplicon detection, it can potentially be used for premortem identification of E. granulosus-infected dogs to enable large-scale surveys in endemic countries where highly specialized equipment to undertake PCR analysis is rare.

摘要

我们之前开发了一种聚合酶链反应(PCR)检测方法,用于检测细粒棘球蚴感染,该方法对鉴定感染犬非常敏感和特异。我们现在开发了一种环介导等温扩增(LAMP)检测方法,该方法可扩增细粒棘球蚴的相同基因组重复序列,用于共检测。该检测方法能够从粪便样本中检测到单个虫卵,并且对细粒棘球蚴具有很高的种特异性,不会与密切相关的寄生虫(包括多房棘球绦虫)的 DNA 发生交叉扩增。由于该方法不需要进行 DNA 扩增的热循环,也不需要电泳检测扩增子,因此它有可能用于生前鉴定感染细粒棘球蚴的犬,从而在高度专业化的 PCR 分析设备稀缺的流行国家进行大规模调查。

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