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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

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【作者】 陈彬王颖郑晶黄晓蓉林杰彭华毅邵碧英

【Author】 CHEN Bin;WANG Ying;ZHENG Jing;HUANG Xiao-rong;LIN Jie;PENG Hua-yi;SHAO Bi-ying;Fujian Entry-Exit Inspection and Quarantine Bureau;Fujian Provincial Key Laboratory of Inspection and Quarantine Technology Research;College of Food Sciences,Fujian Agriculture and Forestry University;

【机构】 福建出入境检验检疫局检验检疫技术中心福建省检验检疫技术研究重点实验室福建农林大学食品科学学院

【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。

【Abstract】 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 100 cfu / m L. 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.

【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)
  • 【文献出处】 食品工业科技 ,Science and Technology of Food Industry , 编辑部邮箱 ,2015年03期
  • 【分类号】TS207.4;O657.72
  • 【下载频次】154
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