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《Fujian Journal of Animal Husbandry and Veterinary》 2007-S1
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Research and application on rapid detection kit of salmonella by universal PCR

Kong Fande1 Chen Qiong2 Xu Shufei1 Peng Haibing3 (1 Xiamen entry-exit inspection and quarantine bureau,Xiamen 361012; 2 Xiamen agricultural product quality and safety testing center,Xiamen 361009; 3 College of Animal science,Fujian agriculture and forestry university,Fuzhou 350002)  
This study evaluated universal PCR to detect Salmonella species according to fimY gene.The specific primers to the fimY gene were used to amplify standard bacteria of A-F salmonella,60 salmonella separate strains and 11 non-salmonella species by PCR. Experiment results indicated that only Salmonella species generated the specific 526 bp DNA fragments on the 2.0% agarose gel and non- salmonella species controls gave negative results,demonstrating that the primers should be specific for Salmonella genus. The PCR was able to detect as little as 93 cfu Salmonella and pre-cultured(BP) or cultured(MM) bacteria liquid of the sample obtaining 3 cfu Salmonella.This indicates established PCR method is high sensitive and strongly specific.The nucleic acid is puried by five methods,including adding bacteria,hot cleavage,freezing-thaw again and again,CTAB and column reagent kit,the CTAB method IS best,but testing procedure is multifarious.Adding bacteria and hot cleavage can procure the same result as column reagent kit and have many merits including simple,rapid,economical and is worthy to generalization and application.
【Fund】: 厦门市科技计划项目(3502Z20042021)资助
【CateGory Index】: S854.43
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