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A gyrB-targeted PCR for Rapid Identification of Salmonella
Authors:Xuhong Ye  Yiming Wang  Xiangui Lin
Affiliation:(1) State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Beijing East Road, 71, Nanjing, 210008, People’s Republic of China;(2) Joint Open Laboratory of Soil and the Environment, Hongkong Baptist University & Institute of Soil Science, Chinese Academy of Sciences, Nanjing, 210008, People’s Republic of China;(3) Graduate University of Chinese Academy of Sciences, Beijing, 100049, People’s Republic of China;(4) Department of Biology and Biochemistry, Institute of Soil Science, Chinese Academy of Sciences, Beijing East Road, 71, Nanjing, 210008, People’s Republic of China;
Abstract:
Salmonella causes the majority of infections in humans and homeothermic animals. This article describes a specific polymerase chain reaction (PCR) method developed for a rapid identification of Salmonella. A gyrB-targeted species-specific primer pair, S-P-for (5′-GGT GGT TTC CGT AAA AGT A-3′) and S-P-rev (5′-GAA TCG CCT GGT TCT TGC-3′), was successfully designed. PCR with all the Salmonella strains produced a 366- bp DNA fragment that was absent from all the non-Salmonella strains tested. The detection limit of the PCR was 0.01 ng with genomic DNA or 3.2 cells per assay. Good specificity was also demonstrated by fecal samples, from which only the gyrB gene of Salmonella was amplified. Using the culture-PCR method, 27 isolates on Salmonella-Shigella (SS) medium were rapidly identified as Salmonella, which was confirmed by the sequencing of the gyrB gene.
Keywords:
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