Wirth D F, Pratt D M
Proc Natl Acad Sci U S A. 1982 Nov;79(22):6999-7003. doi: 10.1073/pnas.79.22.6999.
Kinetoplast DNA (kDNA) was isolated from various species of the protozoic parasite Leishmania and analyzed by nucleic acid hybridization to detect species-related heterogeneity of kDNA. Purified DNA isolated from L. mexicana and L. braziliensis displayed no homology in nucleic acid hybridization studies. These results confirmed that rapid kDNA sequence change and evolution is occurring in New World species of Leishmania and suggested that such isolated kDNA could be used as a specific hybridization probe for the rapid identification of Leishmania species by using whole organisms. This work further demonstrates that such species-specific identification is feasible on isolated Leishmania promastigotes and, more important, directly on tissue touch blots derived from the cutaneous lesion. Thus, specific hybridization of isolated kDNA provides the basis for a rapid, accurate method for the diagnosis of human leishmaniasis directly from infected tissue.
从原生动物寄生虫利什曼原虫的不同物种中分离出线粒体DNA(kDNA),并通过核酸杂交进行分析,以检测kDNA的物种相关异质性。在核酸杂交研究中,从墨西哥利什曼原虫和巴西利什曼原虫分离出的纯化DNA没有显示出同源性。这些结果证实,新世界利什曼原虫物种正在发生快速的kDNA序列变化和进化,并表明这种分离的kDNA可以用作特异性杂交探针,通过使用完整生物体来快速鉴定利什曼原虫物种。这项工作进一步证明,这种物种特异性鉴定在分离的利什曼原虫前鞭毛体上是可行的,更重要的是,直接在源自皮肤病变的组织触片上可行。因此,分离的kDNA的特异性杂交为直接从感染组织中快速、准确地诊断人类利什曼病提供了基础。