Development and Application of the Real-time Recombinase Polymerase Amplification Assay for Detection of Shigella in Food
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    Abstract:

    The aim of the study was to develop a method for the determination of Shigella by real-time RPA. Based on the conserved sequence of invasive plasmid antigen (ipaH) of Shigella, designed specific primer and exo probe, and used to monitor the performance of reaction system by detection equipment, the RPA reaction was performed successfully at 39 ℃ and the results were obtained with 20 min. This method could be specifically detected Shigella, and could not detect non-Shigella bacteria. The study showed that the detection limit of exo-RPA was 1.0×10-3 ng/μL with genomic DNA of Shigella, which was the same as the real-time PCR published articles. For artificially contaminated chicken and broccoli samples with the bacterial concentration of 9 CFU/25 g, the Shigella coule be detected after 8 hours culture by real-time RPA. The result of real-time RPA, which was the same as real-time PCR, but detected need 7-12 min by real-time RPA, and detected at least 35 min by real-time PCR (the Ct was between 27 and 34). The Shigella real-time RPA assay developed in the study was specificity, high sensitivity, simple and rapid, and provided a new technology platform for detection of Shigella.

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  • Online: November 21,2019
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