环介导等温扩增技术快速检测食品中的耐热芽孢菌  

Rapid detection of Thermophilic Bacilli in food by loop-mediated isothermal amplification

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,哈尔滨150030

出　　处：《食品科技》2012年第12期320-323,326,共5页Food Science and Technology

基　　金：十二五国家科技支撑计划项目(2012BAD29B07)

摘　　要：针对食品中的好热黄无氧芽孢菌、嗜热脂肪地芽孢杆菌和凝结芽孢杆菌等耐热芽孢菌,以spo0A基因部分序列为靶基因,设计了一套引物,能够同时对这3种菌进行环介导等温扩增(LAMP)检测,扩增只需要60min。该法快速并具有良好的特异性,灵敏度可达到8cfu/mL,是普通PCR的10倍。DNaseI的使用与LAMP结合,可有效降解死菌游离DNA,消除其影响。结果表明,该方法用于奶粉及罐头制品中耐热芽孢菌的检测,具有灵敏度高、特异性强、检测速度快等优点,在食品微生物检测中拥有广阔的前景。In order to detect Anoxybacillus flavithermus, Geobacillus stearothermophilus, Bacillus coagulans in food, partial spo0A gene sequence was chosen as the target gene, and a set of primers was designed. It takes only 60 min to detect these three bacteria by loop-mediated isothermal amplification （LAMP）. It is a rapid way with good specificity, and its sensitivity can reach 8 cfu/mL, is 10 times as the ordinary PCR. The use of DNase I was connected with LAMP, can effectively degrade the free DNA from dead bacteria. The results showd that this LAMP has proven to be high sensitive, specific and rapid in the detection of Thermophilic Bacillli from milk powder and canned products, and have broad prospects in the detection of food microorqanisms.

关 键 词：好热黄无氧芽孢菌 嗜热脂肪地芽孢杆菌 凝结芽孢杆菌 spo0A基因 环介导等温扩增技术 DNASEI 

分 类 号：TS207.4[轻工技术与工程—食品科学]