环介导等温扩增技术快速检测肉中金黄色葡萄球菌  

Rapid Detection of Staphylococcus aureus in Raw Meat Samples by Loop-mediated Isothermal Amplification

作　　者：赵玥明 满朝新[2] 曲艳艳[1] 姜霞[1] 姜毓君[1,2] 

机构地区：[1]东北农业大学食品学院/乳品科学教育部重点实验室,哈尔滨150030 [2]东北农业大学国家乳业工程技术研究中心,哈尔滨150086

出　　处：《中国食物与营养》2016年第1期57-61,共5页Food and Nutrition in China

基　　金：国家科技支撑计划课题(项目编号:2012BAD28B02;2013BAD18B11;2012BAD29B07)

摘　　要：应用环介导等温扩增（loop-mediated isothermal amplification,LAMP）技术建立了对肉中金黄色葡萄球菌检测的方法。实验中,使用了最新的Bst 2.0 Warm Start DNA聚合酶完成LAMP扩增反应,并针对金黄色葡萄球菌所特有的保守性耐热核酸酶基因（nuc）设计得到了一套LAMP扩增引物。对LAMP法和PCR法的检测灵敏度进行了比较,同时对人工污染肉中的金黄色葡萄球菌进行检测。结果表明：所建立的LAMP法能够特异性的检测金黄色葡萄球菌,并且检测金黄色葡萄球菌纯菌的灵敏度为2.01×10~0CFU/m L,是普通PCR检测灵敏度的100倍。在检测肉中金黄色葡萄球菌时,检测限为2.01×10~1CFU/m L。因此,本实验所建立的LAMP法检测肉中金黄色葡萄球菌的方法,具有灵敏、快速以及简便等的优点,是一种具有很好的发展前景的检测手段。We established a method to detect Staphylococcus aureus in meat by loop-mediated isothermal amplification. During the experiment,using the latest Bst 2. 0 Warm Start DNA polymerase to finish the LAMP amplification reaction,and against S. aureus-specific nuclease resistant conserved gene（ nuc） designed LAMP amplification primers. the detection sensitivity of LAMP and the PCR method were compured,while was detect Staphylococcus aureus. the artificially contaminated meat The results showed that the built LAMP method could specifically detect Staphylococcus aureus,and Staphylococcus aureus bacteria in pure sensitivity 2. 01 × 100 CFU / m L,which was 10~0 times the normal PCR detection sensitivity. the detection limit of detecting Staphylococcus aureus in meat was 2. 01 × 10~1 CFU / m L. Thus,the LAMP method to detect Staphylococcus aureus in meat was established,which had the advantage of sensitive,rapid and simple,which was a kind of good prospects for development of testing methods.

关 键 词：环介导等温扩增 金黄色葡萄球菌 肉 

分 类 号：R155.55[医药卫生—营养与食品卫生学]