中文摘要：

目的：建立环介导等温扩增（LAMP）检测大肠埃希菌耐药基因ampC的方法，并评价其临床应用价值。方法：设计3对特异性LAMP引物，建立25μL的LAMP检测体系并进行优化，确定最佳反应时间；与PCR进行比较检测其特异性和灵敏度：对54株临床分离的大肠埃希菌分别进行LAMP检测和药敏试验，评价检测耐药基因的临床应用价值。结果：成功建立了检测大肠埃希茵耐药基因ampC的LAMP方法，在65℃保温1h可实现对ampC基因的快速检测，其特异性与PCR一致，灵敏度高出PCR约10倍，耐药基因的检测与药敏试验有统计学差异（P=0.000）。结论：LAMP检测技术可望发展为快速、准确检测细菌耐药基因的方法，但目前尚不能替代药敏试验指导临床用药。

英文摘要：

Objective To establish a method of loop-mediated isothermal amplification （LAMP） for detection of ampC gene from E.coli and evaluate its application value in clinic. Methods 3 pairs of primers were designed, used to establish a 25 μL LAMP reaction system. The LAMP reaction mix and the best reaction time were optimized, its sensitivity and specificity were evaluated by comparing with PCR. 54 strains of E.coli from clinic were detected by LAMP and drug sensitive test respectively, analyzed the results with the statistic methods to assess the value of LAMP in clinical application. Results The LAMP reaction system was successfully established and optimized, it could correctly identify the ampC gene when incubated at 65 ℃ for 60 min. LAMP had the same specificity with PCR, but its sensitivity was 10 times higher. There were significant differences between LAMP and drug sensitive test （P = 0.000）. Conclusions LAMP is promising to be developed into an effective method for detecting bacterial drug resistance gene rapidly and accurately, but at present it can＇t replace the drug sensitive test for guiding the use of medicines.