水生生物学报2017，Vol.41Issue(6)：1225-1231,7.DOI:10.7541/2017.152
翘嘴鳜病原嗜水气单胞菌分子特征及LAMP检测方法的建立
MOLECULAR CHARACTERIZATION AND ESTABLISHMENT OF LAMP DETECTION METHOD OF PATHOGENIC AEROMONAS HYDROPHILA ISOLATED FROM SINIPERCA CHUATSI
摘要
Abstract
Aeromonas hydrophila exists in the water body,which can cause many kinds of diseases and huge loss of aquaculture animals to the aquaculture production.The pathogenicity and virulence characteristics of the isolated strain which caused mass mortalities of reared Mandarin fish were studied.The rapid detection methods ofA.hydrophila were established by the loop-mediated isothermal amplification (LAMP) using aerA gene as molecular marker.The results showed that the LDs0 ofA.hydrophila was 1.6× 106 CFU/mL,and 14 virulence genes were detected from the isolated strain,which can be biology markers in detecting of pathogenic A.hydrophila.The assay of LAMP for detecting A.hydrophila was established using aerA gene as molecular marker.Positive reactions were detected by the stair-step amplified bands,and the green amplified products were observed directly by naked-eye in the reaction tube by addition of SYBR Green Ⅰ.No any amplified bands and orange color were detected in control group.The results showed that LAMP detection method ofA.hydrophila had good specificity.The results of sensitivity of LAMP showed that the primers could detect A.hydrophila at the level of4.6×l01 CFU/mL.The artificially infected 10 species of freshwater fishes and shrimps revealed positive amplified results using LAMP detection method,and no-infected 10 samples revealed negative amplified results.The results showed that the LAMP detection method has preferable application,and could be used in rapid diagnose of aquatic animal diseases caused by A.hydrophila.关键词
翘嘴鳜/嗜水气单胞菌/aerA基因/环介导等温扩增(LAMP)Key words
Siniperca chuatsi/Aeromonas hydrophila/aerA gene/Loop-mediated isothermal amplification (LAMP)分类
农业科技引用本文复制引用
张悦,高晓建,叶金明,陈楠,杜雪地,张晓君,邴旭文..翘嘴鳜病原嗜水气单胞菌分子特征及LAMP检测方法的建立[J].水生生物学报,2017,41(6):1225-1231,7.基金项目
江苏省水产三项工程项目(Y2016-33和D2015-15) （Y2016-33和D2015-15）
江苏省高校自然科学研究重大项目(14KJA240001) （14KJA240001）
江苏省重点研发计划项目(BE2017311)资助[Supported by the Projects of Shui Chan San Xiang of Jiangsu Province (Y2016-33,D2015-15) （BE2017311）
the Major Natural Science Research Program of Jiangsu Higher Education Institutions (14KJA240001) （14KJA240001）
the Key Research and Development Program of Jiangsu Province (BE2017311)] （BE2017311）
