应用环介导等温扩增技术对分离自呼吸系统感染铜绿假单胞菌的快速检测

Establishment and optimization of loop-mediated isothermal amplification for rapid detection of pseudomonas aeruginosa

目的:建立环介导恒温扩增技术(LAMP)快速检测铜绿假单胞杆菌的方法。方法选取分离自呼吸系统感染的铜绿假单胞杆菌152例,针对铜绿假单胞菌菌种特异性基因ETA建立LAMP扩增反应,研究反应灵敏度、特异性以及实际应用。结果:PCR法所能检测到的最低DNA量为132 pg,而LAMP法则为132 fg,LAMP法比PCR法的灵敏度高1 000倍。LAMP法对标准菌株以及分离自呼吸系统感染的152株铜绿假单胞菌,检出率与特异性分别达94.7%与100%。结论:LAMP是一种可应用于对呼吸系统感染铜绿假单胞菌的灵敏、特异快速检测方法。

Objective To establish a Loop-mediated Isothermal Amplification（LAMP） for Pseudomonas aeruginosa rapid detection. Method 152 P. aeruginosa strains isolated from nasal swabs and 30 reference strains were applied. P. aeruginosa ATCC15442 was used to develop LAMP amplification and evaluate sensitivity and specificity. Results Sensitivity of LAMP was 103 times higher than PCR, with DNA amount as 132 fg. When LAMP was applied to 30 reference strains and 152 P. aeruginosa strains, the specification was 100% while identification rate reached 94.7%. Conclusion The establishment LAMP showed a promising prospect in P. aeruginosa rapid detection.