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目的： 分析我国不同类型流行区内脏利什曼原虫 （Ｌ．ｄ．） 分离株ｋＤＮＡ。方法： 应用利什曼原虫属特异引物１３Ａ， １３Ｂ及据杜氏利什曼原虫四川人分离株ｋＤＮＡ 微环特异片段序列设计合成的引物Ⅰ、引物Ⅱ， ＰＣＲ扩增不同流行区利什曼原虫分离株ｋＤＮＡ， 获特定片段， 进行ＳＳＣＰ分析。结果： 用引物Ⅰ和引物ⅡＰＣＲ扩增内脏利什曼原虫分离株ｋＤＮＡ， 在同样试验条件下， 山丘地区和荒漠地区Ｌ．ｄ． 分离株扩增出２９７ ｂｐ 特定片段， 而平原地区Ｌ．ｄ．分离株及新疆皮肤利什曼原虫未扩增出２９７ ｂｐ 特定片段。将上述各虫株的２９７ ｂｐ 特定片段进行ＳＳＣＰ分析， 可见两个山丘地区的Ｌ．ｄ．分离株ｓｓＤＮＡ 迁移率相同， 而与荒漠地区新疆７７１分离株则相差较大。用引物１３Ａ， １３ＢＰＣＲ 扩增平原地区的Ｌ．ｄ．山东分离株和Ｌ．ｄ．江苏分离株、山丘地区的Ｌ．ｄ．汶川分离株、Ｌ．ｄ．甘肃分离株，均扩增出１２０ ｂｐ 特定片段。经ＳＳＣＰ分析，平原地区的Ｌ．ｄ．山东分离株和Ｌ．ｄ．江苏分离株的ｓｓＤＮＡ迁移率完全相同； 山丘地区的Ｌ．ｄ．汶川分离株和Ｌ．ｄ． 甘肃分离株ｓｓＤＮＡ迁移率相同， 但与平原地区者明显不同； 婴儿利什曼ｓｓＤＮＡ迁移
Objective: To analyze kdDNA of L.d. isolates from different types of endemic areas in China. Methods: Primers I and II were designed and synthesized by using Leishmania-specific primers 13A and 13B and kDNA-specific fragment sequences of Leishmania donovani isolates, and PCR amplification of Leishmania isolates from different endemic areas Strains of kDNA were spotted for SSCP analysis. Results: The karyotype of Leishmania isolates kDNA was amplified by PCR with primer Ⅰ and primer Ⅱ. In the same experimental conditions, the hill area and the desert area L. d. The isolate amplified 297 bp specific fragment, while the plains L. d. The specific fragment of 297 bp was not amplified from the isolate and Leishmania cutata. SSCP analysis of the 297 bp specific fragments from each of the above mentioned isolates revealed L in two hills. d. The isolates had the same mobility of ssDNA, while the isolates of 771 in desert region of Xinjiang were quite different. Primers 13A, 13BPCR were used to amplify L in the plains. d. Shandong isolate and L. d. Jiangsu isolates, L. in the hills. d. Wenchuan isolate, L. d. Gansu isolates, were amplified 120 bp specific fragments. After SSCP analysis, L in the plains. d. Shandong isolate and L. d. Jiangsu isolates ssDNA mobility exactly the same; hills L. d. Wenchuan isolate and L. d. Gansu isolate ssDNA mobility of the same, but significantly different from the plain areas; infant Leishman’s ssDNA migration