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目的确定rDNA的内转录间隔区Ⅰ(ITS-1)是否可作为我国杜氏利什曼原虫(Leishmania donovani)分类的分子遗传标记。方法对我国不同疫区杜氏利什曼原虫分离株(L.d.SD1、L.d.SC7、L.d.XJ3、L.d.JS12)的ITS-1进行PCR扩增、克隆、测序,并分析它们的ITS-1序列多态性。结果 4个不同疫区的利什曼原虫分离株(L.d.SD1、L.d.SC7、L.d.XJ3、L.d.JS12)经PCR均扩增出一条约320bp的ITS-1片段,其序列大小分别为:317bp、323bp、317bp、316bp。序列分析表明:序列之间在多处位点发生了不同类型的突变,存在明显的序列多态性,其中伽师县突变的频率最为频繁,序列多态性最为显著。结论 ITS-1可以作为Leishmania donovani种内分类的分子遗传标记,有利于我国内脏利什慢原虫病的防治、诊断和治疗。
Objective To determine whether the internal transcribed spacer region (ITS-1) of rDNA can be used as a molecular genetic marker for Leishmania donovani classification in China. Methods ITS-1 of Leishmania donovani isolates (LdSD1, LdSC7, LdXJ3, LdJS12) from different epidemic areas of China were amplified by PCR, cloned and sequenced, and their ITS-1 sequence polymorphisms . Results A total of 320 bp ITS-1 fragment was amplified by PCR from four Leishmania isolates (LdSD1, LdSC7, LdXJ3 and LdJS12) from different epidemic areas. The sequence size was 317 bp and 323 bp, respectively , 317 bp, 316 bp. Sequence analysis showed that there were different types of mutations at different sites between sequences, and obvious sequence polymorphism existed. Among them, Jiashi County had the most frequent mutation and the most significant sequence polymorphism. Conclusion ITS-1 can be used as a molecular genetic marker for intraspecific taxonomic classification of Leishmania donovani, which is beneficial to the prevention, diagnosis and treatment of visceral Leishmaniasis in China.