欢迎您！东篱公司
退出
-
申报数据库
-
立项数据库
-
成果数据库
-
项目获奖数据库
中文摘要：
针对鲫鱼诺卡氏菌16S-23SrRNA基因内转录间隔序列，设计了四条LAMP引物，在外内引物浓度比1：8、dNTP浓度0．8—1．2mmol／L、Mg2．浓度10--14mmol／L、反应温度60—65℃、反应时间60min的优化条件下，扩增产物经电泳后呈现特异性的LAMP梯形条带，建立的LAMP法具有高灵敏度，达到10^-7μg／μlDNA浓度，比常规PCR方法高100倍。将该方法应用于取自养殖场的乌鳢、大黄鱼、黄姑鱼组织样品检测，结果在16份组织样品中有7份检出自然感染的鳓鱼诺卡氏菌，与同步取样品鱼组织的细菌分离、培养检查结果相一致，并显示既能检测发病鱼，又能检出未发病且已感染的病鱼。分析表明，这是一种能快速、简易、特异、敏感的检测鱼类致病鲫鱼诺卡氏菌的诊断方法。
英文摘要：
To develop a loop-mediated isothermal amplification （LAMP） assay for detection of fish pathogenic Nocardia seriolae. Four primers were designed based on the sequence of the 16S--23S ribosomal RNA internal transcribed spacer region of N. seriolae for the use of LAMP assay. The assay was optimized to the following conditions： 1：8 outer primer concentration ratio, 0.8--1.2mmol/L of dNTP, 10--14mmol/L of Mg2＋, at 60--65℃ for 60min. LAMP amplification products had ladder-like bands when electrophoresed on an agarose gel. The assay could amplify up to 10^-7μg/μl of DNA. Hence, the sensitivity of LAMP assay was 100-fold higher than the standard PCR protocol. The assay was applied to detect the presence of Nocardia seriolae in different tissue samples from farm Ophiocephalus argus, Pseudnoseiaena crotea, Niber albiflora. Seven naturally infected 17. seriolae were detected in 16 tissue samples, and the results from the bacterial culture test were consistent with fish tissue samples taken simultaneously. This study showed LAMP assay could be used not only to diagnose the infected fishes, but also to recognize the carrier ofN. seriolae as well. LAMP assay is a rapid, simple, specific and sensitive test to diagnose for fish pathogenic N. seriolae.
同期刊论文项目
同项目期刊论文
期刊信息
位置：