FOOD SCIENCE ›› 2019, Vol. 40 ›› Issue (2): 298-303.doi: 10.7506/spkx1002-6630-20170906-094

• Safety Detection • Previous Articles     Next Articles

Development and Application of a Recombinase Polymerase Amplification Assay for Detection of Salmonella

LIU Libing1,2, GENG Yunyun3, JIANG Yanfen1, LIU Siying3, SUN Xiaoxia1,2, NAN Huizhu1, WANG Jianchang1,2,*   

  1. (1. Center of Inspection and Quarantine, Hebei Entry-Exit Inspection and Quarantine Bureau, Shijiazhuang 050051, China; 2. Hebei Academy of Inspection and Quarantine, Shijiazhuang 050051, China; 3. College of Life Science, Hebei Normal University Hebei, Shijiazhuang 050024, China)
  • Online:2019-01-25 Published:2019-01-22

Abstract: The aim of the study was to develop a method for the determination of Salmonella by recombinase polymerase amplification (RPA). A specific primer pair was designed based on the conserved sequence of the invasion protein A gene (invA) of Salmonella. The reaction time was optimized and the RPA method was performed successfully at 38 ℃ for 20 min in a water bath: the target fragment was effectively amplified. The RPA reaction could specifically detect Salmonella rather than 26 other foodborne pathogens. The detection limit (LOD) of RPA was 1.1 × 10-3 ng/μL with the genomic DNA of Salmonella as a template, which was consistent with that of real-time PCR. For artificially contaminated lamp, chicken and broccoli samples with a bacterial concentration of 4 CFU/25 g, Salmonella could be detected by RPA after 8 hours of culture. Consistent results were obtained using real-time PCR. The RPA assay was specific, simple and rapid, and could represent a new direction for the detection of foodborne pathogens.

Key words: Salmonella, invA, recombinase polymerase amplification (RPA), detection

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