Amplification of small subunit rRNA sequences of trypanosomatids from human samples of a patient diagnosed with visceral leishmaniasis.. ssrRNA sequences of trypanosomatids in human samples.

Tissue samples from a patient diagnosed with visceral leishmaniasis in northeast region of Brazil were used for DNA isolation in order to detect trypanosomatid small subunit RNA sequence. We used DNA samples from liver,spleen, blood, bone marrow and skin biopsy in PCR using primers described by Noyes et al (International Journal for Parasitology v. 29, p. 331-339, 1999). Amplicons were sequenced by Sanger method and phylogenetic analysis showed that the parasites were more related to other Leishmaniinae parasites, such as Crithidia and Leptomonas (one-host parasites exclusive for insects), rather than Leishmania species.

本研究采集巴西东北部地区一名确诊为内脏利什曼病（visceral leishmaniasis）患者的组织样本，用于DNA提取以检测锥虫科（trypanosomatid）小亚基RNA序列。实验采用肝脏、脾脏、血液、骨髓及皮肤活检的DNA样本，使用Noyes等人于1999年发表于《International Journal for Parasitology》第29卷第331-339页所描述的引物开展聚合酶链式反应（PCR）扩增。扩增产物（amplicons）通过桑格（Sanger）法完成测序，系统发育分析结果显示，该寄生虫与利什曼亚科（Leishmaniinae）的其他类群（如专性寄生昆虫的单宿主寄生虫克里格锥虫属（Crithidia）和细滴虫属（Leptomonas））的亲缘关系，相较于利什曼原虫属（Leishmania）物种更为密切。