我国山丘疫区和平原疫区利什曼原虫分离株基因组DNA基因型分析

ANALYSIS OF NUCLEAR DNA GENE TYPES OF LEISHMANIA ISOLATES FROM HILLY AND PLAIN FOCI OF CHINA

目的：分析我国山丘和平原疫区利什曼原虫分离株基因组ＤＮＡ（ｎＤＮＡ）的多态性。方法：对基因组ＤＮＡ进行内切酶酶切、Ｓｏｕｔｈｅｒｎ杂交、染色体定位。探针采用地高辛标记。结果：采用ｇｐ６３探针，显示杜氏利什曼原虫（Ｌｅｉｓｈｍａｎｉａｄｏｎｏｖａｎｉ，Ｌ．ｄ．）江苏人株和Ｌ．ｄ．ＪｅｄｄａｈｎＤＮＡ之间有相似的染色体杂交带，Ｌ．ｄ．四川人株和Ｌ．ｉｎｆａｎｔｕｍｎＤＮＡ之间有相似的染色体杂交带；采用β－ｔｕｂｕｌｉｎ探针，显示Ｌ．ｄ．江苏人株与Ｌ．ｄ．ＪｅｄｄａｈｎＤＮＡ之间有两条相似的染色体杂交带，Ｌ．ｄ．四川犬株与Ｌ．ｄ．甘肃犬株ｎＤＮＡ之间有三条相似的染色体杂交带、与Ｌ．ｄ．汶川人株ｎＤＮＡ之间有两条相似的染色体杂交带。结论：提示平原疫区的江苏人株与Ｌ．ｄ．Ｊｅｄｄａｈ之间存在同源性，山丘疫区的Ｌ．ｄ．四川人株与Ｌ．ｉｎｆａｎｔｕｍ之间存在同源性，Ｌ．ｄ．四川犬株与Ｌ．ｄ．甘肃犬株之间存在同源性、与Ｌ．ｄ．汶川人株之间既存在同源性又存在差异。

AIM: To analyse the nuclear DNA (nDNA) polymorphism of Leishmania isolates from hilly and plain foci of China. METHODS: nDNA were analysed by endonuclease digestion, Southern blotting and chromosomal localization. Probes were labeled with digoxigenin. RESULTS: Using gp63 gene probe, similar hybridization bands were found to be existed between nDNAs of Leishmania donovani Jiangsu human isolate and L.d.Jeddah, also between nDNAs of L.d. Sichuan human isolate and L.infantum. Using β tubulin gene probe,there were two similar hybridization bands existed between nDNAs of L.d.Jiangsu human isolate and L.d.Jeddah, and three similar hybridization bands existed between nDNAs of L.d.Sichuan canine isolate and L.d. Gansu canine isolate, and two similar hybridization bands existed between nDNA of L.d. Sichuan canine isolate and L.d. Wenchuan human isolate.CONCLUSION: Homology exists between L.d. Jiangsu human isolate and L.d. Jeddah from plain foci, between L.d. Sichuan human isolate from hilly foci and L.infantum, between L.d.Sichuan canine isolate and L.d. Gansu canine isolate from hilly foci. Homology as well as differences exist between L.d. Sichuan canine isolate and L.d. Wenchuan human isolate. Heterogeneity exists between isolates from hilly foci and plain foci.