Rapid detection of Salmonella by enzymatic recombinase amplification combined with lateral flow chromatography
NIE Yanni1,2, YAN Meiying3, SONG Yanyan2
1. Baotou Medical College, Inner Mongolia University of Science and Technology, Baotou, Inner Mongolia 014040, China; 2. Department of Microbiological Testing, Chaoyang District Center for Disease Control and Prevention, Beijing 100020, China; 3. Chinese Center for Disease Control and Prevention, Beijing 102206, China
Abstract：Objective To establish a rapid detection method for Salmonella based on the combination of enzymatic recombinase amplification (ERA) and lateral flow chromatography (LF), so as to provide technical support for the on-site detection of Salmonella. Methods Specific ERA primers and probes were designed based on the highly conserved flagella gene fimY in Salmonella. The primers were screened using capillary electrophoresis, and the probes were designed according to the amplification range of the screened primers. The amplification temperature and time were optimized to establish the amplification method, and the product was detected using LF strips. A standard strain of Salmonella was used to verify the sensitivity, 10 other gut bacteria were used to to verify the specificity and sensitivity, and the nucleic acid of the actual Salmonella strains was amplified to verify the detectability. Results After screening for Salmonella-specific primers using capillary electrophoresis, the minimum detection concentration was 5 copies/μL under the amplification temperature of 37 ℃ and reaction time of 20 minutes. This method had a positive amplification result for Salmonella nucleic acid, and the amplification results of 10 other gut bacteria were all negative, with good specificity. Conclusion This method provides a possibility for on-site point of care testing of Salmonella infection.
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