LAMP法在阪崎肠杆菌快速检测中的应用

The application of loop mediated isothermal amplification(LAMP)technique for rapid detection of Ent erob a cter sakazakii

针对阪崎肠杆菌的ompA基因设计4条特异性引物(2条内引物、2条外引物),建立一种快速检测阪崎肠杆菌的DNA环介导恒温扩增法(LAMP)。通过对2株阪崎肠杆菌标准菌株、24株阪崎肠杆菌分离株和18株非阪崎肠杆菌测试,结果表明此方法具有很高的特异性;对其灵敏度进行检验,发现该方法的最低检测限可以达到8CFU/mL,具有较好的敏感性。在1份实验室能力验证盲样和26份实际奶粉样品中应用发现,该方法与传统生化方法结果吻合,具有较好的可靠性。

Four primers including two inner primers,two outer prim-ers were designed specifically to recognize the ompA gene of E.sakazakii and a method of loop-mediated isothermal amplification was set up to detect Enterobacter sakazakii rapidly.The results sug-gested that the method had high speciality to 2 E.sakazakii typical strains,24isolates and 18non-E.sakazakii.More over,it had good sensitivity with the lowest detectable limit of 8CFU/mL.Through detecting the proficiency tests of E.sakazakii in milk pow-der and 26milk powder samples by this method,the results indicated that it had the same results as traditional biochemical method and may be used to detect samples.