荧光定量PCR快速检测食品中肠出血性大肠杆菌O104

目的建立针对肠出血性大肠杆菌O104的荧光定量PCR快速检测方法。方法针对O104的wzx保守基因,设计特异性引物和探针,采用倍比梯度稀释法检测该体系的灵敏度,以另外34株肠道致病菌评价检测方法的特异性;建立了肠出血性大肠杆菌O104感染食品的检测模型以验证方法的适用性。结果建立了荧光定量PCR快速检测肠出血性大肠杆菌O104的方法。每次检测最低限为12.0copies,特异性为100%。结论该法缩短了检测时间,并有良好的灵敏性和特异性,在疾病防控及食品卫生行业中具有应用前景。

Rapid detection of enterohemorrhagic escherichia coli O104 in food with Real-time PCR assay

Objective To develop a real-time PCR assay for rapid detection of enterohemorrhagic Escherichia coli O104.Methods The specific primers and probes were designed to amplify the wzx gene of EHEC O104.And the sensitivity of system was detected by using fold dilution method.In order to examine the specificity of the system,other 34 intestinal bacteria strains were assayed and mimic EHEC O104 infected food was detected for evaluation.Results A highly sensitive and specific real-time PCR assay was established for detection of EHEC O104.The sensitivity was 12 copies/ test for EHEC O104.The specificity was 100%.Conclusion The real-time PCR detection method is efficient to detect EHEC O104 with good sensitivity and specificity.The method has good prospects of application in disease prevention and food safety area.