二重PCR法快速检测3种食源性致病菌

目的建立快速检测食品中金黄色葡萄球菌(Staphylococcus aureus)、沙门氏菌属(Salmonella spp.)及志贺氏菌属(Shigella spp.)的二重PCR方法。方法分别针对金黄色葡萄球菌.femA和nuc、沙门氏菌属invA和hilA及志贺氏菌属ipaB和ipaH设计6对特异性引物,检测每对引物的特异性及灵敏度,组合优化各自二重PCR反应体系。结果初步建立了针对这3类致病菌的二重PCR检测方法,整个检测过程不超过18 h,检测灵敏度均可达10 pg DNA/reaction。结论所建立的针对3类致病菌的二重PCR检测方法具有灵敏度高、特异性强和方便高效等优点,具有良好的市场应用前景。

Duplex polymerase chain reaction methods for rapid determination of 3 foodborne bacterial pathogens

To construction the multiplex PCR systems for rapid detection foodborne bacterial pathogens Staphylococcus aureus, Salmonella spp. and Shigella spp. in food, respectively. Method Six pairs of specific primers were designed according to the specific genes femA and nuc of Staphylococcus aureus, genes invA and hilA of Salmonella spp., and ipaB and ipaH of Shigella spp.. Compared the detection sensitivity of each pair of primers, and then associated and optimized the multiplex PCR reaction conditions and systems. Results All these multiplex PCR detection systems were constructed with many advantages, including high sensitivity, strong specificity, convenient and efficient to operation; and the whole detection can be performed in less than 15 h with all detection sensitivities reach 10 pg. Conclusion These multiplex PCR detection systems and methods provided a key technical support for developing kits for foodborne bacterial pathogens rapid detection, and have favorable applications perspective.