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

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.