【摘要】 目的 探讨中华血吸虫（Schistosoma sinensium, S. sinensium）与日本血吸虫（Schistosoma japonicum, S. japonicum）种间亲缘性，为寻找2种血吸虫的差异性基因片段提供参考；同时探讨日本血吸虫成虫与尾蚴之间的基因变异，为日本血吸虫不同生活史阶段的分子检测方法建立提供参考。方法 通过随机扩增多态DNA(random amplified polymorphic DNA, RAPD)技术，利用40个随机引物分别对日本血吸虫尾蚴、成虫，中华血吸虫尾蚴和成虫样本DNA进行3次重复PCR扩增实验。根据扩增结果中1个扩增重复性、稳定性较好的样本，统计有效随机引物扩增条带数；计算日本血吸虫与中华血吸虫的相似系数（genetic similarity, S）和遗传距离（genetic distance, D），分别分析日本血吸虫与中华血吸虫种间差异和日本血吸虫种内DNA多态性差异；运用SPSS 26.0进行聚类分析。结果 筛选出29个有效随机引物，扩增出日本血吸虫共173个条带，多态性条带占45.66%(79/173)；中华血吸虫共332个条带，多态性条带占68.37%(227/332)。种间分析结果显示，二者尾蚴分别扩增出83个、128个条带，有35个共同条带，尾蚴间S为0.33,D为0.67；成虫分别扩增出90个、96个条带，共同条带为33个条带，成虫间S为0.35,D为0.65；日本血吸虫种内分析结果显示，A县1村尾蚴和成虫分别扩增出83个、90个条带，有48个共同条带，S为0.55,D为0.45。聚类结果显示，所有样本可分为日本血吸虫和中华血吸虫2大支，日本血吸虫支又可分为尾蚴支和成虫支。中华血吸虫尾蚴支按县（市）划分C县尾蚴支和X县尾蚴支。成虫支同样按县（市）划分X县成虫支和C县成虫支。结论 日本血吸虫与中华血吸虫在尾蚴和成虫间均有相似扩增带型，二者存在一定的亲缘关系，可根据随机引物S35、P256、S07扩增出的条带进一步查找2种血吸虫的差异性基因；日本血吸虫尾蚴与成虫基因间存在变异，在建立血吸虫分子检测技术时对于采用尾蚴和成虫作为检测样本应分别进行充分验证。更多还原

【Abstract】 ObjectiveTo explore the interspecific affinity between Schistosoma sinensium and Schistosoma japonicum,and to provide a reference for identifying differential gene fragments between the two species. Additionally, this study aimed to assess the genetic variation between the adult worms and cercaria of S. japonicum, offering insights for developing molecular detection methods tailored for different life history stages of S. japonicum.MethodsUsing the random amplified polymorphic DNA(RAPD) technique, DNA samples from cercariae and adults of S. japonicum and S. sinensium were subjected to triplicate PCR amplification with 40 random primers. Based on the amplification results, one sample with good amplification repeatability and stability was selected for further analysis. The number of amplified bands from effective random primers was counted, and genetic similarity(S) and genetic distance(D) between S. japonicum and S. sinensium were calculated to evaluate the interspecific differences, as well as intraspecific DNA polymorphism within S. japonicum. Cluster analysis was performed using SPSS 26.0.ResultsTwenty-nine effective random primers were screened out, yielding 173 bands for S. japonicum, of which45.66%(79/173) were polymorphic bands; for S. sinensium, 332 bands were amplified, with 68.37%(227/332) being polymorphic bands. Interspecific analysis revealed that the cercariae of S. japonicum and S. sinensium produced 83 and 128 bands, respectively, with 35 bands in common, resulting in a genetic similarity(S) of 0.33 and a genetic distance(D) of 0.67;adults produced 90 and 96 bands, respectively, with 33 common bands, showing S=0.35 and D=0.65. Intraspecific analysis of S. japonicum from Village 1, County A, showed that cercariae and adult worms yielded 83 and 90 bands, respectively, sharing48 common bands(S=0.55, D=0.45). Cluster analysis results showed that all samples could be divided into two major branches:S. japonicum and S. sinensium. The S. japonicum branch could be further divided into the cercariae branch and the adult branch. The S. sinensium cercariae branch was divided by county(or city) into County C and County X sub-branches, while the adult branch was similarly divided into County X and County C sub-branches.Conclusion S. japonicum and S. sinensium have similar amplification band patterns in both cercarial and adult stages, indicating a certain affinity between them. Bands amplified by random primers S35, P256, and S07 can be used to further identify the differential genes between the two species.Genetic variations exist between the cercarial and adult stages of S. japonicum; therefore, the existence of genetic variation between the cercariae and adult stages of S. japonicum underscores the need for separate validation of molecular detection assays for each life stage.更多还原