基于微卫星标志构建湖北钉螺PCR多重反应体系  

Study on A Multiplex PCR System of Oncomelania Hupensis Based on Microsatellites

作　　者：周支军 沈璐[1] 华雪涛 顾漫漫 邹惠莹 吕大兵[2] ZHOU Zhijun;SHEN Lu;HUA Xuetao;GU Manman;ZOU Huiying;LU Dabing(Wuzhong District Center for Disease Control and Prevention,Suzhou 215128,Jiangsu Provice,China;Department of Epidemiology and Health Statistics,School of Public Health,Soochow University,Suzhou 215123,Jiangsu Province,China)

机构地区：[1]苏州市吴中区疾病预防控制中心,江苏苏州215128 [2]苏州大学公共卫生学院流行病与卫生统计学系,江苏苏州215123

出　　处：《寄生虫病与感染性疾病》2021年第3期163-167,共5页Parasitoses and Infectious Diseases

基　　金：苏州市2019年度第十八批科技发展项目(民生科技)资助(项目编号:SS2019044);国家自然科学基金(项目编号:81971957)。

摘　　要：目的筛选湖北钉螺多态性微卫星位点,建立多重反应体系,以对钉螺进行基因分型和群体遗传学分析。方法从GenBank数据库和文献中筛选合适的湖北钉螺微卫星位点,设计合成引物并进行评价,构建多重反应体系。结果从GenBank中获得4个满足条件的三核苷酸重复序列位点,从文献中获得9个双核苷酸序列位点;成功设计合成11对引物,其中5对引物扩增效果好。最终构建的多重反应体系中5个位点均具有高度多态性,且不存在连锁不平衡。结论研究所建立的多重反应体系扩增效果好、各位点多态性高,将有利于后续开展针对钉螺扩散的溯源研究。Objective To screen the polymorphic microsatellite loci of Oncomelania hupensis,and to establish a multiple reaction system,so that to monitor and trace the spread of O.hupensis.Methods Appropriate microsatellite loci of O.hupensis were screened from the public GenBank and from the published literatures.The corresponding primers were then designed and synthesized.Highly polymorphic loci were chosen and finally a multiplex PCR system were developed.The polymorphism of each locus were assessed.Results Four loci with trinucleotide repeat sequences were acquired from the GenBank and nine loci with dinucleotide repeat sequences from literatures.Eleven pairs of primers were successfully designed and synthesized,of which 5 pairs of primers had good amplification effects.A multiplex PCR system with five highly polymorphic loci was developed.All five loci each were highly polymorphic,and there was no linkage disequilibrium between them.Conclusion The multiplex PCR system containing five high polymorphic microsatellite loci will facilitate the follow-up research on monitoring the spread of snails.

关 键 词：湖北钉螺 微卫星 多态性 多重反应体系 

分 类 号：R184[医药卫生—流行病学] R122.2[医药卫生—公共卫生与预防医学]