餐饮食品中金黄色葡萄球菌的实时荧光PCR检测方法

Detection of Staphylococcus Aureus in Catering Food by Real-Time Polymerase Chain Reaction

文中研发一种快速检测餐饮食品中金黄色葡萄球菌的方法。研究方法采用煮沸裂解法提取样品DNA,运用实时荧光PCR法检测样品中的金黄色葡萄球菌特异性基因片段。文中比较了不同的DNA制备方法的优劣,根据金黄色葡萄球菌基因组中的保守序列设计特异性引物,建立了快速检测餐饮食品中金黄色葡萄球菌的方法,并对阳性培养液及餐饮食品中的金黄色葡萄球菌进行检测。结果表明,采用文中方法与引物可在7份餐饮食品中扩增出金黄色葡萄球菌特异性的基因片段。实时PCR检测可检出浓度低至1.0×10^(-5)μg/μL的金黄色葡萄球菌DNA,检测时间少于24h。文中方法可以广泛应用于餐饮食品等食品中金黄色葡萄球菌的高通量快速筛查检测,具有高效、快速、灵敏、准确等优点。

The objective of this study is to validate an easy-to-use Pre-PCR sample preparation method based on a simple boiling protocol for screening of Staphylococcus aureus in catering food using PCR method.According to the conserved area of Staphylococcus aureus genome,specific primers were designed to detect Staphylococcus aureus in catering food by using PCR techniques.Specific sequences were amplified by PCR,which is matched to the genomic data about Staphylococcus aureus published on GeneBank.To the real-time PCR,the sensitivity can detect 1.0×10^-5μg/μL Staphylococcus aureus DNA and the whole detection time was less than 24h.In general,the PCR method established in this study was a kind of sensitive and specific detection technique for Staphylococcus aureus in catering food.