三种利什曼原虫和我国新疆皮肤利什曼原虫kDNA分子杂交实验研究

Study on the Molecular Hybridization in kDNA of 3 Leishmania Species and the Pathogenic Agent of Cutaneous Leishmaniasis in Xin-jiang, China

本研究自新疆克拉玛依地区皮肤利什曼病（CL）患者的皮肤病变组织内抽提微量的利什曼原虫kDNA，采用引物13A、13B进行PCR扩增，获120bP扩增区带（CL－PCR－AP），并转移到尼龙膜上，分别与地高辛（Digoxigenin－11－dUTP）标记的L．tropica、L．infantum、L．gerbillikDNA探针进行Southern印迹杂交。结果：病变组织内利什曼原虫kDNA的PCR扩增区带仅与LtropicakDNA探针有明显的杂交信号，而与L.infantum、L.gerbillikDNA探针杂交则呈阴性反应。结论：LtropicakDNA与新疆克拉玛依地区CL患者的CL－PCR－AP之间存在同源关系。

in this paper, a 120hp kDNA fragments of Leishmania from cutaneous lesions of 2 parasitologically confirmed patients and 3 Leishmania species (L. tropica L. infantum and L. gerbilli )were directly amplified with primer 13A and 13B. Southern hybridization were conducted with digoxigenin labelled L. tropica, L. infantum, L. gerbilli kDNA probes respectively. Results showed thatstrong hybridization was found to occur only in CL-PCR-AP with L. tropica kDNA probe, while L. infantum and L. gerbilli id)NA probes showed no detectable signals. Our results confirmed that homologous sequences exists in kDNA of L. tropica and the species causing CL in. Xinjiang, China.