二重PCR法快速检测3种食源性致病菌

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

中文关键词:  二重PCR  金黄色葡萄球菌  沙门氏菌属  志贺氏菌属  快速检测

英文关键词:duplex PCR  Staphylococcus aureus  Salmonella spp.  Shigella spp.  rapid detection

中文摘要:

      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。

英文摘要:

      Objective To establish a duplex PCR system for rapid detecting foodborne bacterial pathogens such as 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 genes ipaB and ipaH of Shigella spp. respectively. The corresponding detection specificity and sensitivity of each pair of primers were measured, and each duplex PCR reaction systems were combinatorial optimized. Results The duplex PCR detection systems for rapid detecting the above-mentioned foodborne bacterial pathogens were preliminary established, the whole detection was performed in less than 18 h with all of the detection sensitivities reached 10 pg DNA/reaction. Conclusion The duplex PCR detection systems showed many advantages, including with high sensitivity and strong specificity, convenient and efficient to operate. So it has a prospective application in market.