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Visual detection technique for efficient screening and isolation of <Emphasis Type="Italic">Salmonella</Emphasis> based on a novel enrichment assay using chromatography membrane
Authors:F Tang  Y Xiong  H Zhang  K Wu  Y Xiang  J-B Shao  H-W Ai  Y-P Xiang  X-L Zheng  J-R Lv  H Sun  L-S Bao  Z Zhang  H-B Hu  J-Y Zhang  L Chen  J Lu  W-Y Liu  H Mei  Y Ma  C-F Xu  A-Y Fang  M Gu  C-Y Xu  Y Chen  Z Chen  Z-Y Sun
Institution:1.Department of Laboratory Medicine, Tongji Medical College,Tongji Hospital, Huazhong University of Science and Technology,Wuhan,People’s Republic of China;2.Department of Laboratory Medicine,Wuhan Children’s Hospital, Huazhong University of Science and Technology,Wuhan,People’s Republic of China;3.Microbiological Laboratory, Wuhan Center for Disease Control and Prevention,Wuhan,People’s Republic of China;4.Microbiological Laboratory, Qiaokou Center for Disease Control and Prevention,Wuhan,People’s Republic of China;5.Jiangan Center for Disease Control and Prevention,Wuhan,People’s Republic of China;6.Department of CT/MRI,Wuhan Children’s Hospital, Huazhong University of Science and Technology,Wuhan,People’s Republic of China;7.Department of Rehabilitation, Tongji Medical College,Tongji Hospital, Huazhong University of Science and Technology,Wuhan,People’s Republic of China;8.Clinic Department of Hong Kong Road,Wuhan Children’s Hospital, Huazhong University of Science and Technology,Wuhan,People’s Republic of China;9.Department of Research and Education,Wuhan Children’s Hospital, Huazhong University of Science and Technology,Wuhan,People’s Republic of China;10.Department of Transfusion Service,Wuhan Medical and Health Centre for Women and Children,Wuhan,People’s Republic of China;11.Microbiological laboratory, Jianghan Center for Disease Control and Prevention,Wuhan,People’s Republic of China;12.Department of Gastroenterology,Wuhan Children’s Hospital, Huazhong University of Science and Technology,Wuhan,People’s Republic of China;13.Hubei Provincial Institute for Food Supervision and Test,Wuhan,People’s Republic of China;14.Department of Laboratory Medicine,Sixth People’s Hospital of Wuhan,Wuhan,People’s Republic of China;15.Department of Pharmacy, Tongji Medical College,Xiehe Hospital, Huazhong University of Science and Technology,Wuhan,People’s Republic of China;16.Department of Laboratory Medicine,Wuhan Hospital of Traditional Chinese Medicine,Wuhan,People’s Republic of China;17.Department of Gastroenterology,Pu Ai Hospital of Huazhong Technology and Science University,Wuhan,People’s Republic of China
Abstract:To detect Salmonella more efficiently and isolate strains more easily, a novel and simple detection method that uses an enrichment assay and two chromogenic reactions on a chromatography membrane was developed. Grade 3 chromatography paper is used as functionalized solid phase support (SPS), which contains specially optimized medium. One reaction for screening is based on the sulfate-reducing capacity of Salmonella. Hydrogen sulfide (H2S) generated by Salmonella reacts with ammonium ferric citrate to produce black colored ferrous sulfide. Another reaction is based on Salmonella C8 esterase that is unique for Enterobacteriaceae except Serratia and interacts with 4-methylumbelliferyl caprylate (MUCAP) to produce fluorescent umbelliferone, which is visible under ultraviolet light. A very low detection limit (101 CFU ml?1) for Salmonella was achieved on the background of 105 CFU ml?1 Escherichia coli. More importantly, testing with more than 1,000 anal samples indicated that our method has a high positive detection rate and is relatively low cost, compared with the traditional culture-based method. It took only 1 day for the preliminary screening and 2 days to efficiently isolate the Salmonella cells, indicating that the new assay is specific, rapid, and simple for Salmonella detection. In contrast to the traditional culture-based method, this method can be easily used to screen and isolate targeted strains with the naked eye. The results of quantitative and comparative experiments showed that the visual detection technique is an efficient alternative method for the screening of Salmonella spp. in many applications of large-sized samples related to public health surveillance.
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