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目的从基因组序列中选择和设定新的、可用于研究日本血吸虫多态性的微卫星位点。方法672个日本血吸虫标本取自安徽、江西、湖南、湖北和四川等5省的7个流行区现场。根据日本血吸虫基因组数据库中的初始数据设计引物,对标本的17个全新的日本血吸虫微卫星位点进行分析。用GenMapper4.0软件(Applied Biosystems)进行初始数据处理,用GenClone1.0软件计算种群内各标本的遗传距离;用GenAlEx6软件统计等位基因的变化;用GenClone1.0软件计算种群内各标本的遗传距离;用UPGMA谱系图分析种群间遗传距离。结果17个微卫星位点均呈现多态性。对江西都昌种群的这17个位点进行分析,标本之间的遗传距离大多为25～32,显示江西都昌种群内各个标本之间的遗传差异明显。对7个地区种群分析的结果显示种群间的差异较为明显。类平均法系统聚类分析(UPGMA)谱系图显示,江西都昌、安徽铜陵、湖北沙市和湖南常德的种群同为一簇,其遗传距离为0.0178～0.0363;安徽贵池和湖南岳阳的为另外一簇,其遗传距离为0.0247;而四川西昌单独为一簇,与其他两簇的遗传距离为0.0192～0.0693。结论选择的17个微卫星位点可作为研究日本血吸虫种群遗传学的有效标记,并揭示中国大陆该虫种存在着复杂的遗传变异。
Objective To select and set up new microsatellite loci which can be used to study the polymorphisms of Schistosoma japonicum from genome sequences. Methods A total of 672 schistosoma japonicum samples were collected from 7 endemic areas in 5 provinces including Anhui, Jiangxi, Hunan, Hubei and Sichuan. According to the initial data of Schistosoma japonicum genome database, primers were designed and 17 new S. japonicum microsatellite loci were analyzed. The initial data was processed with GenBiosper4.0 software (Applied Biosystems), the genetic distance of each specimen in the population was calculated by GenClone1.0 software, the allele was analyzed by GenAlEx6 software, and the genetics of each specimen in the population was calculated by GenClone1.0 software Distance; UPGMA pedigree analysis of genetic distance between populations. Results 17 microsatellite loci showed polymorphism. The 17 sites of Duchang population in Jiangxi Province were analyzed. The genetic distance between the two species was mostly 25-32, indicating that the genetic differences among all the samples in Duchang were significant. The results of population analysis in seven regions showed that the differences among the populations were more obvious. Cluster analysis of UPGMA showed that the populations of Duchang in Anhui, Tongling in Anhui, Shashi in Hubei and Changde in Hunan were clustered together and the genetic distance was 0.0178 ~ 0.0363. The genetic diversity of Guichi in Anhui and Yueyang in Hunan was different A cluster with a genetic distance of 0.0247, while Xichang, Sichuan alone was a cluster with a genetic distance of 0.0192-0.0693 to the other two clusters. Conclusion The 17 selected microsatellite loci can be used as effective markers for studying the population genetics of Schistosoma japonicum and reveal the complex genetic variation of this species in mainland China.