金黄色葡萄球菌五种肠毒素基因PCR-DHPLC检测方法的建立  

Development of PCR- DHPLC detection method for five types of Staphylococcal enterotoxin genes

作　　者：陈彬[1,2] 王颖[1,3] 郑晶[1,2] 黄晓蓉[1,2] 林杰[1,2] 彭华毅[1,2] 邵碧英[1,2] 

机构地区：[1]福建出入境检验检疫局检验检疫技术中心,福建福州350001 [2]福建省检验检疫技术研究重点实验室,福建福州350001 [3]福建农林大学食品科学学院,福建福州350002

出　　处：《食品工业科技》2015年第3期172-177,共6页Science and Technology of Food Industry

基　　金：福建省科技厅自然科学基金项目(2012J01062);福建局科技项目(FK2012-25)

摘　　要：本文设计合成了5种肠毒素基因的特异性引物,对PCR的退火温度、DNA模板量进行优化,建立了5种肠毒素基因(SEA、SEB、SEC、SED、SEE)的聚合酶链式反应-变性高效液相色谱(PCR-DHPLC)检测方法,并测定方法的灵敏度和特异性。结果表明建立的PCR-DHPLC检测方法的特异性很好,灵敏度可达到100cfu/m L。PCR-DHPLC方法具有快速、准确、高通量等优点,在进出口食品中金黄色葡萄球菌肠毒素的检测上有很好的应用价值。Five pairs of primers were designed and synthesized for five Staphylococcal enterotoxins genes （SEA, SEB,SEC,SED, SEE）. The PCR-DHPLC detection methods for five Staphylococcal enterotoxins genes were established after optimizing of the annealing temperature and DNA template amount.The sensitivity and specificity of the detection method were then determined.The results showed that the PCR- DHPLC method could detect specifically of Staphylococcal enterotoxins genes with detection limit of 100cfu/mL. The PCR- DHPLC detection method had some advantages,such as rapid,accuracy and high throughput.The PCR-DHPLC method had fine application value to detecting Staphylococcal enterotoxins in import and export foods.

关 键 词：金黄色葡萄球菌 肠毒素 聚合酶链式反应(PCR) 变性高效液相色谱(DHPLC) 

分 类 号：TS201.3[轻工技术与工程—食品科学]