[目的]初步探讨常规PCR、荧光PCR、PCR产物测序及脉冲场凝胶电泳(PFGE)等核酸检测技术在常见细菌性食源疾病病原菌检测中的应用。[方法]用常规PCR、荧光PCR直接检测经选择性增菌培养的沙门菌和志贺菌,测定PCR扩增产物序列以确定病原菌种类,用限制性内切酶XbaI和SpeI对分离菌株进行PFGE分析。[结果]常规PCR、荧光PCR方法在两份腹泻病人样本中均检出沙门菌,结果与培养法一致。二者未在皮蛋中检出沙门菌,而培养法在市售皮蛋中检出沙门菌。检测时间分别为PCR16h、荧光PCR14h、PCR产物直接测序19h,较培养法缩短4d。来源不同的3株分离菌XbaI和SpeI酶切图谱完全一致(相似度为100%)。[结论]应用常规PCR、荧光PCR可准确、及时检出腹泻病人样本中的沙门菌和志贺菌,有望用于食物中毒的快速检测。但在检测食品和环境样本时需适当延长增菌培养时间。在从农田到餐桌的食品供应链中,PFGE可对疾病暴发进行早期识别并协助对病原菌及其传播途径的溯源,有助于食物中毒的预防。 [Objective] The research aimed to discuss the application of routine PCR, fluorescent PCR, PCR product sequencing and pulsed-field gel electrophoresis (PFGE) in the detection of common bacterial pathogenic bacteria. [Method] Salmonella and Shigella were selectively detected by conventional PCR and fluorescent PCR. The PCR products were sequenced to determine the pathogen species. The isolates were analyzed by PFGE with restriction endonucleases XbaI and SpeI . [Result] Salmonella was detected in both samples of diarrhea by conventional PCR and fluorescence PCR, and the results were consistent with the culture method. The two did not detect Salmonella in preserved eggs, while culture method detected Salmonella in commercially available preserved eggs. The detection time was PCR16h, fluorescence PCR14h, PCR products were sequenced directly 19h, compared with the culture method shortened 4d. Three isolates from different sources XbaI and SpeI digestion maps exactly the same (similarity of 100%). [Conclusion] Salmonella and Shigella can be detected accurately and timely by routine PCR and fluorescence PCR in diarrhea patients, which is expected to be used for the rapid detection of food poisoning. However, in the detection of food and environmental samples need to be extended lengthening culture time. In farm-to-table food supply chains, PFGE can early identify disease outbreaks and help trace sources of pathogens and their routes of transmission, helping to prevent food poisoning.