剪股颖粒线虫环介导等温扩增检测技术研究  

Detection of Anguina agrostis by loop-mediated isothermal amplification method

作　　者：俞禄珍 张博文 焦彬彬 戚龙君 田沂民 宋绍禕 Yu Luzhen;Zhang Bowen;Jiao Binbin;Qi Longjun;Tian Yimin;Song Shaoyi(Shanghai Customs District,Shanghai 200135,China)

机构地区：[1]上海海关

出　　处：《植物检疫》2019年第5期27-31,共5页Plant Quarantine

基　　金：国家重点研发计划(2017YFC1200600);原上海出入境检验检疫科研项目(HK003-2017)

摘　　要：剪股颖粒线虫(Anguina agrostis)是牧草上重要的病原线虫,被我国列入检疫性有害生物名单。本研究根据剪股颖粒线虫的rDNA-ITS区序列设计特异性的引物和探针,建立了环介导等温扩增和核酸试纸条检测技术。结果表明,该LAMP-核酸试纸条检测法的特异性强,供试2个剪股颖粒线虫虫株均为阳性,其他23种线虫均为阴性;灵敏度高,最低检测限为3.15ng,检测灵敏度达1/400条2龄幼虫。该法操作简单,对设备要求低,适合口岸推广应用。Anguina agrostis is an important pathogenic nematode on pasture and is listed as quarantine pest of China.Primers and probes set were designed based on rDNA-ITS region sequences to develop a loop-mediated isothermal amplification (LAMP) and nucleic acid strip method for A.agrostis detection.It showed that the method had advantages of good specificity:positive reactions were observed in the presence of A.agrostis,whereas all other 23 species nematodes showed negative reactions.The detection limit of the method was 3.15 ng,and it could detect 1/400 nematode of a single 2nd larva.Thus,this method has advantages of easy operation and low requirements of expensive equipment.It is suitable for applying in basic laboratory.

关 键 词：剪股颖粒线虫 环介导等温扩增 核酸试纸条 检测 

分 类 号：S41-30[农业科学—植物保护]