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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。
OBJECTIVE To establish a rapid real-time fluorescence quantitative PCR for detection of enterohemorrhagic Escherichia coli O104. Methods According to the conserved gene wzx of O104, specific primers and probes were designed. The sensitivity of the system was tested by fold gradient dilution method. The specificity of the method was evaluated by 34 strains of enteropathogenic bacteria. The intestinal hemorrhagic large intestine Bacillus O104 infects the food testing model to verify the suitability of the method. Results The method of rapid detection of enterohemorrhagic Escherichia coli O104 by fluorescence quantitative PCR was established. Each test has a minimum of 12.0 copies and a specificity of 100%. Conclusion This method has shortened the detection time, has good sensitivity and specificity, and has application prospect in the field of disease prevention and control and food hygiene.