LAMP在乳品厂耐热菌检测中的应用  

Application of LAMP for rapid detection of thermophilic bacilli in dairy factory

作　　者：窦明理[1] 霍贵成[1] 

机构地区：[1]东北农业大学乳品科学教育部重点实验室,黑龙江哈尔滨150030

出　　处：《食品与机械》2012年第5期75-78,196,共5页Food and Machinery

基　　金："十二五"国家科技支撑计划(编号:2012BAD29B07)

摘　　要：好热黄无氧芽孢菌和凝结芽孢杆菌是乳品中常见的污染菌。为了检测该两种菌,以spo0A基因片段为靶基因,通过序列比对,设计出环介导等温扩增技术(LAMP)引物,对spo0A基因部分序列进行了扩增,并对反应温度、甜菜碱浓度、dNTPs浓度及Mg2+浓度等扩增条件进行优化。结果表明:在温度63℃,甜菜碱浓度0.6 mol/L,dNTPs浓度1.0mmol/L、Mg2+浓度6.0mmol/L时,反应体系孵育30～60min,扩增效果最佳,检测灵敏度能够达到51CFU/mL。Anoxybacillus flavithermus and Bacillus coagulans are common pollutant bacteria in milk powder.In order to detect these two kinds of bacteria,spo0Agenes were aligned and chosed as target gene,and primers were designed for loop-mediated isothermal amplification（LAMP）.Also the amplification conditions including reaction temperature,betaine,dNTPs and Mg2＋concentrations were optimized.It amplified best at 63 ℃,betaine concentration at 0.6mol/L,dNTPs concentration at 1.0mmol/L,Mg2＋concentration at 6.0mmol/L,incubation 30～60min.The sensitivity of the LAMP assay was 51CFU/mL.

关 键 词：好热黄无氧芽孢菌 凝结芽孢杆菌 环介导等温扩增技术 快速检测 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]