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| Establishment of a recombinase polymerase amplification fluorescence detection method for Yersinia pestis |
| PEI Meijuan1,2,3, LI Shulei1,2,3, LI Menglan1, LIU Chaoyang1, SONG Na1, WANG Yufei1 |
1. Department of Laboratory Medicine, the Third Medical Centre, Chinese PLA General Hospital, Beijing 100039,China;
2. Jinzhou Medical University, Jinzhou 121001,China;
3. Institute of Bioengineering, Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100071,China |
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Abstract Objective To establish a simple, rapid method of the recombinase polymerase amplification (RPA) technology for the detection of Yersinia pestis.Methods Primers and probes were designed basing on specific sequences of Yersinia, and the reaction temperature was optimized by detecting different temperatures. The specificity of this method was evaluated by detecting DNA templates of different strains, and the sensitivity was determined by testing sample DNA templates of different dilution concentrations. The application of this method was evaluated by simulating samples.Results Specific primers and probe were designed based on the DNA adenine methylase gene and the fluorescence RPA method was established, which can successfully detect up to 100 copies of Yersinia pestis gene in 20 minutes at 35 ℃, and no cross-reaction with other bacteria, the simulation of the sample detection accuracy of 98.0%.Conclusions RPA could be used in the clinical detection of Yersinia pestis, especially under crude conditions and the method is helpful to the prevention and control of plague.
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Received: 15 November 2021
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2022, Vol. 33 
