嗜肺军团菌核酸等温扩增快速检测技术研究  

Research of rapid isothermal nucleic acid amplification testing technology for Legionella pneumoila

作　　者：张兵 徐昌平[2] 程东庆[1] 冯燕[2] 卢亦愚[2] ZHANG Bing;XU Chang- ping;CHENG Dong- qing;FENG Yan;LU Yi -yu(Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310053, China)

机构地区：[1]浙江中医药大学,浙江杭州310053 [2]浙江省疾病预防控制中心,浙江杭州310009

出　　处：《中国卫生检验杂志》2018年第7期782-784,788,共4页Chinese Journal of Health Laboratory Technology

基　　金：浙江省医学重点学科群建设项目(XKQ-009-003);浙江省卫生适宜技术成果转化计划(2018ZH007);免疫金渗滤法快速检测军团菌(2016R410044)

摘　　要：目的建立一种嗜肺军团菌的核酸等温扩增试纸条及pH显色快速检测方法,从而实现对空调冷却水样中嗜肺军团菌的快速现场检测。方法基于嗜肺军团菌特异性mip基因设计引物,建立环介导等温扩增体系,并结合反应产物横向流动试纸条和pH显色检测技术,进而验证方法的灵敏性、特异性,并对浙江省不同地区的中央空调冷却水水样进行检测。结果环介导的等温扩增试纸条及pH显色检测方法灵敏度可高达3 cfu/μl,特异性好,且检测时间仅需40 min左右。检测的灵敏度与荧光定量PCR的方法基本一致,对外环境空调冷却水的检测阳性率与荧光定量也相符。结论建立的嗜肺军团菌试纸条与pH显色快速检测方法,灵敏度高,特异性好,适合现场快速检测。Objective To establish an isothermal nucleic acid amplification dipsticks and pH color rapid detection method for Legionella pneumoila,so as to realize rapid field test of Legionella pneumoila in the air conditioning cooling water samples.Methods Design primers based on Legionella pneumoila specificity mip gene,build loop-mediated isothermal amplification system,and to combine the reaction product lateral flow dipstick with pH color detection technology,and then verify sensitivity and specificity of the method.The samples in central air conditioning cooling water was enrolled into test.Results The sensitivity of this method can reached 3 cfu/μl,the specificity was good,and testing time was only about 40 minutes.The sensitivity of the detection was basically as same as the method of fluorescence quantitative PCR.The positive rate of external environmental air conditioning cooling water detection was also consistent with fluorescence quantification.Conclusion The established lateral flow dipstick and pH color rapid detection method has the advantages of high sensitivity,good specificity,which is suitable for on-site rapid detection.

关 键 词：嗜肺军团菌 MIP基因 环介导的等温扩增技术 横向流动试纸条 pH显色 

分 类 号：R511.5[医药卫生—内科学]