日本血吸虫尾蚴高表达基因LAMP检测方法初探  

Detection of Highly Expressed Genes in Schitosoma Japonicum Cercaria by Using Loop-mediated isothermal Amplification:a Preliminary Study

作　　者：杨树国 张轶静 关玮 李健 赵燕清 

机构地区：[1]湖北医药学院基础医学院寄生虫学教研室,湖北十堰442000

出　　处：《湖北医药学院学报》2017年第4期304-308,F0003,共6页Journal of Hubei University of Medicine

基　　金：湖北省卫生计生血防专项(WJ2015XB031);湖北省教育厅指导项目(B2015475)

摘　　要：目的:应用LAMP方法对日本血吸虫尾蚴阶段高表达基因簇进行初步检测,为群体的血吸虫早期预警提供参考。方法:选择日本血吸虫尾蚴期高表达基因,利用在线软件Primer Explorer V4设计目的基因引物。纯水室温孵育感染性钉螺,收集尾蚴,并提取尾蚴和钉螺基因组DNA,用LAMP方法检测尾蚴和感染性钉螺期日本血吸虫的基因表达情况。结果:在所选择的16个基因中,CNUS0000102858.1、CNUS0000103598.1、CNUS0000103614.1等8个蛋白编码基因的LAMP检测尾蚴DNA结果为阳性;CNUS0000038.1、CNUS0000102858.1、CNUS0000103614.1等8个蛋白编码基因的LAMP扩增感染性钉螺DNA结果为阳性。其中,LAMP检测尾蚴及感染性钉螺基因组DNA结果均为阳性的基因有CNUS0000102858.1、CNUS0000103598.1、CNUS0000103614.1、CNUS0000105798.1、CNUS0000106268.1、CNUS0000106483.1、CNUS0000107429.1。结论:在16个备选基因中,这7个尾蚴及感染性钉螺基因组DNA检测结果均为阳性的基因,可考虑作为钉螺及疫水样本LAMP检测基因,为日本血吸虫流行病学调查和群体早期预警提供实验数据。Objective Detected the genes coding high expression proteins by using Loop-mediated isothermal amplification in Schistosoma japonicum cercaria and infective oncomelania genomic DNA. Methods The high expression protein genes were selected and primers were designed using online software Primer Explorer V4. The infective oncomelanias were incubated in ultrapure water and cercarias were collected. The cercaria and infective oncomelania genomic DNA were extracted and the positive genes were screened by using Loop-mediated isothermal amplification. Results 16 selective genes were tested in cercaria genomic DNA, and the positive reactions were happened in 8 genes including CNUS0000102858. 1、CNUS0000103598.1、CNUS0000103614.1. The same positive reactions were watched in 8 genes exist in infective oncomelania genomic DNA including CNUS0000038.1、CNUS0000102858.1、CNUS0000103614.1. Among these,CNUS0000102858.1、CNUS0000103598. 1、CNUS0000103614. 1、CNUS0000105798. 1、CNUS0000106268. 1、CNUS0000106483. 1、CNUS0000107429.1 were detected positive by using LAMP which happened in cercaria and infective oncomelania genomic DNA. Conclusion These 7 genes could be used to the oncomelania and infected water LAMP detecting and epidemiological investigation of Schistosoma japonicum.

关 键 词：日本血吸虫 尾蚴 感染性钉螺 环介导扩增技术 

分 类 号：R383.24[医药卫生—医学寄生虫学]