摘要
利用环介导等温扩增技术(LAMP)建立了一种对嗜水气单胞菌(Aeromonas hydrophila)进行快速检测的方法。基于嗜水气单胞菌tbp A基因,设计了一套引物,并优化了反应体系的温度、时间和Mg2+浓度,分析了该方法的特异性与灵敏度。结果表明,在59℃条件下,LAMP的最适扩增时间为40 min,能有效检测到1 pg/μL核酸浓度,且检测出嗜水气单胞菌为阳性,其他菌株为阴性,该方法耗时短,灵敏度高,特异性好,适合嗜水气单胞菌的快速检测。
LAMP（Loop-mediated isothermal amplification） as a kind of rapid detection method for Aeromonas hydrophila was established. A set of primers were designed based on the sequences of the tbp A gene of A. hydrophila. Temperature, time,Mg2＋concentration of reaction system were optimized, and detected the specificity and sensitivity of this method. The results showed that under 59 ℃, the LAMP amplification optimum time for 40 min, this method could effectively detect the nucleic acid concentration with 1 pg/μL, and the LAMP of A. hydrophila reactions was positive, others were negative. In a conclusion, the LAMP detective method was short time-consuming, sensitivity and specificity; and it was suitable for rapid detection of A. hydrophila.
出处
《湖北农业科学》
2015年第20期5125-5128,共4页
Hubei Agricultural Sciences
基金
国家自然科学基金青年项目(31402341)
博士后科学基金面上项目(2014M550400)
淡水生态与生物技术国家重点实验室开放基金项目(2015FB05)
珠江所农业部渔用药物创制重点实验室开放基金项目(201306)
