溶血性链球菌LAMP检测方法的建立  

Establishment of Loop-mediated Isothermal Amplification for the Detection of Streptococcus hemolyticus

作　　者：尹欢[1] 李琦[1] 陈江源[1] 马寅众[1] 李睿[1] 刘志国[1] 

机构地区：[1]武汉工业学院生物与制药工程学院,湖北武汉430023

出　　处：《食品科学》2010年第22期311-314,共4页Food Science

基　　金：武汉市科技局对外科技合作与交流计划项目(201070934341);武汉工业学院研究生创新基金重点项目(09cx002)

摘　　要：目的:研究溶血性链球菌的环介导等温扩增(loop-mediated isothermal amplification,LAMP)检测方法,实现对溶血性链球菌的快速检测。方法:针对溶血性链球菌的scpA基因设计4条特异性LAMP内外引物进行LAMP扩增;优化扩增反应条件;采用包括溶血性链球菌在内的6种不同菌株进行LAMP的特异性检测,并酶切鉴定;对溶血性链球菌菌液以无菌水进行10倍系列梯度稀释后,进行LAMP扩增检测其灵敏度;将菌掺入牛奶样本并进行LAMP检测。结果:本法可快速灵敏地检测出溶血性链球菌,反应特异性高,检出限为16.7CFU/mL,而牛奶样本的检出限则达10CFU/mL。结论:LAMP法可用于食品溶血性链球菌的快速检测。Objective： To develop a loop-mediated isothermal amplification （LAMP） method for the rapid detection of Streptococcus hemolyticus. Methods： According to the scpA gene of Streptococcus hemolyticus, 4 specific primers were designed for LAMP amplification. Six different Streptococcus hemolyticus strains were validated by LAMP and identified by enzymatic digestion. The sensitivity of LAMP method was evaluated by serial gradient dilution of Streptococcus hemolyticus solution. Results： LAMP could be used for the rapid and sensitive detection of Streptococcus hemolyticus with a high sensitivity and a detection limit of 16.7 CFU/mL. Moreover, the detection limit was 10 CFU/mL in milk samples. Conclusion： LAMP method is a simple, rapid and feasible method for the detection of Streptococcus hemolyticus.

关 键 词：溶血性链球菌 环介导等温扩增(LAMP) 检测 检出限 

分 类 号：Q933[生物学—微生物学]