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目的了解中国荒漠、山丘和平原疫区利什曼原虫分离株的种群遗传学和流行病学特点。方法选用7个微卫星标记(Lm2TG,Li21-34,Li71-19,Li45-24,Li71-7,Li71-33,Li72-17/2)分别对中国不同疫区的5株杜氏利什曼原虫分离株(9044、SC9、771、LIU和XU)与WHO杜氏利什曼原虫参照株(MHOM/IN/80/DD8)进行PCR扩增和测序,分析序列的微卫星多态性,使用MEGA5.0软件构建系统发育树,推断杜氏利什曼原虫的种群结构。结果 6株杜氏利什曼原虫分离株均扩增出7个微卫星位点。7个位点在杜氏利什曼原虫中检出3~7个等位基因,基因多样性最低为0.523 8(Li45-24),最高为1.000(Li72-17/2)。构建的系统发育树中,中国不同疫区的5株分离株与WHO参照株聚合为一群,其中山丘疫区四川犬分离株表现出明显的多态性。结论中国不同疫区杜氏利什曼原虫分离株的微卫星序列存在丰富的多态性,种系发育分析显示利什曼原虫遗传多态性与地理来源之间存在相关性。
Objective To understand the population genetic and epidemiological characteristics of Leishmania isolates in the desert, hills and plains of China. Methods Seven microsatellite markers (Lm2TG, Li21-34, Li71-19, Li45-24, Li71-7, Li71-33 and Li72-17 / 2) were used to detect Leishmania donovani The microsatellite polymorphisms of the sequences (9044, SC9,771, LIU and XU) and WHO Leishmania donovani reference strain (MHOM / IN / 80 / DD8) were analyzed by PCR using MEGA5. 0 Software to construct phylogenetic tree to infer the population structure of Leishmania donovani. Results Six microsatellite loci were amplified from six strains of Leishmania donovani. Seven loci detected Leishmania donovani 3-7 alleles, the lowest genetic diversity was 0.523 8 (Li45-24), the highest was 1.000 (Li72-17 / 2). Among the phylogenetic trees constructed, five isolates from different epidemic areas in China aggregated with WHO reference strains, and the canine isolates from Sichuan Province showed obvious polymorphism. Conclusion There are abundant polymorphisms in the microsatellite sequences of Leishmania donovani isolates from different epidemic areas in China. Phylogenetic analysis shows that Leishmania genetic polymorphism is related to geographical origin.