Objective To detect enterohemorrhagic Escherichia coli(EHEC)O157∶H7,visualization recombinant enzyme polymerase amplification(RPA)detection and real-time fluorescence RPA detection methods were established based on RPA technology.Methods Primers and probes were designed based on the rfbE gene of EHEC O157∶H7,and specificity and sensitivity were determined.A real-time fluorescent RPA method was established.Combined with the color change characteristics of SYBR Green Ⅰ in nucleic acid reactions,a visualization RPA method for the detection of EHEC O157∶H7 was established.Results The established real-time fluorescence RPA detection and visual RPA detection methods both detected EHEC O157∶H7 in 25 min at a reaction temperature of 35 ℃.The designed primers and probes showed specific results only for EHEC O157∶H7.The detection limits of the two methods were lower than 10-5 ng/μL.Conclusion The visual RPA detection and real-time fluorescence RPA detection methods of EHEC O157∶H7 were easy to operate with strong specificity and high sensitivity.This study provides novel methods for the rapid detection of EHEC O157∶H7 in the field and laboratory.
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