多重PCR检测4种常见食源性致病菌

目的 建立一种多重聚合酶链式反应法(multiplex polymerase chain reaction, MPCR)快速检测肉制品中金黄色葡萄球菌、沙门氏菌、志贺氏菌和单增李斯特氏菌的分析方法。方法 选取金黄色葡萄球菌nuc基因、沙门氏菌SipB基因、志贺氏菌ipaH基因、单增李斯特菌inlA基因作为目标基因, 设计4对PCR引物, 建立并优化多重PCR反应体系, 评价该体系的特异性和灵敏度, 并对人工污染的熟肉样品进行检测。结果 构建的多重PCR方法特异性强、灵敏度高, 人工污染熟肉匀浆中4种致病菌的检出限为103 CFU/mL。结论 构建的多重PCR检测方法能够快速、准确、高效地检测肉制品中金黄色葡萄球菌、沙门氏菌、志贺氏菌和单增李斯特氏菌, 为食源性疾病菌的快速检测提供参考依据。

Detection of four common foodborne pathogens by multiplex PCR

Objective To establish a method for simultaneous and rapid determination of Staphylococcus aureus, Salmonella spp., Shigella spp. and Listeria monocytogenes in food by multiplex polymerase chain reaction (MPCR). Methods Four pairs of primers were designed based on nuc gene for Staphylococcus aureus, SipB gene for Salmonella spp., ipaH gene for Shigella spp., inlA gene for Listeria monocytogenes. After developing and optimizing the multiplex PCR reaction system, the specificity and sensitivity of the multiple PCR assay was evaluated, and then the system was applied in the artificially contaminated cooked meat for the detection of the 4 food-borne pathogens. Results The multiplex PCR method established in this experiment had strong specificity and high sensitively, the limit of detection of the 4 food-borne bacterial pathogens above-mentioned in artificially contaminated cooked meat was 103 CFU/mL. Conclusion The established multiplex PCR method is rapid, sensitive and specific for the detection of Staphylococcus aureus, Salmonella spp., Shigella spp. and Listeria monocytogenes, and can lay the basis for rapid the detection of food-borne pathogens.