摘要
DNA环介导等恒扩增(loop-mediated isothermal amplification,LAMP)技术是一种特异、灵敏、快速的新型基因检测技术,其检测结果可以通过加入核酸燃料SYBR GreenⅠ进行肉眼观察,也可以通过琼脂糖凝胶电泳进行观察。本文以结核分支杆菌(M.TB)作为研究对象,设计了4种LAMP引物以特异地识别结核分支杆菌中gyrB基因的六个特殊区域,着重于利用LAMP技术在几种常见的致病分支杆菌中能快速、特异地检测出结核分支杆菌。实验结果表明,LAMP技术能在恒温(63℃)条件下,1h内检测出结核分枝杆菌,分别对7株非结核分枝杆菌进行检测,只有一株偶发分支杆菌得到了阳性的结果。说明该技术有望应用于畜牧业中肉制品或乳制品中结核分支杆菌的检测。
Loop-mediated isothermal amplification（LAMP） method is a novel gene detection technique with specificity, sensitive and rapidity, the products of which can be visually inspected by adding nucleic acid dye（SYBR Green I ）, or electrophoresed in 2% agarose gels. In this paper, the application of LAMP in the rapid detection of Mycobacterium Tuberculosis was researched. A set of four LAMP primers were designed for recognizing six distinct regions on the target DNA （gyrB）. The results showed that Mycobacterium Tuberculosis was rapidly and specifically detected by LAMP method among several common isolates within 1 h at constant temperature of 63 ~C. The specificity of the method was validated on 7strains of nontubercnlosis mycobacterial with only one strain showing positive result. So the LAMP method will be used for detection of Mycobacterium Tuberculosis in meat or milk products in animal husbandry for food safety.
出处
《现代食品科技》
EI
CAS
2008年第8期835-838,共4页
Modern Food Science and Technology
基金
2005年粤港关键领域重点突破项目(2005A11601101)
