Visualization and real-time fluorescence RPA detection methods for enterohemorrhagic Escherichia coli O157:H7
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1.Qingdao International Travel Health Care Center, Shandong Qingdao 266000, China;2.Huangdao Customs Port Administration of the People’s Republic of China, Shandong Qingdao 266000, China

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R155

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    Abstract:

    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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XU Ying, ZHUANG Guodong, WANG Lili, TENG Xindong. Visualization and real-time fluorescence RPA detection methods for enterohemorrhagic Escherichia coli O157:H7[J].中国食品卫生杂志,2024,36(1):26-31.

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  • Received:November 14,2022
  • Revised:
  • Adopted:
  • Online: April 09,2024
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