Establishment and Application of a Visual DNA Microarray for the Detection of Food-borne Pathogens
文献类型： 外文期刊
作者： Li, Yongjin 1 ;
作者机构： 1.Huzhou Univ, Chinese Acad Fishery Sci, Key Lab Aquat Anim Genet Breeding & Nutr, Zhejiang Prov Key Lab Aquat Resources Conservat &, Huzhou 313000, Peoples R China
2.Yangtze Univ, Coll Life Sci, Jingzhou 434025, Peoples R China
关键词： Micro-plate chip;food-borne pathogen;visual detection;multiplex PCR
期刊名称：ANALYTICAL SCIENCES （ 影响因子：2.081； 五年影响因子：1.676 ）
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收录情况： SCI
摘要： The accurate detection and identification of food-borne pathogenic microorganisms is critical for food safety nowadays. In the present work, a visual DNA microarray was established and applied to detect pathogens commonly found in food, including Salmonella enterica, Shigella flexneri, E. coli O157:H7 and Listeria monocytogenes in food samples. Multiplex PCR (mPCR) was employed to simultaneously amplify specific gene fragments, fimY for Salmonella, ipaH for Shigella, iap for L. monocytogenes and ECs2841 for E. coli O157:H7, respectively. Biotinylated PCR amplicons annealed to the microarray probes were then reacted with a streptavidin-alkaline phosphatase conjugate and nitro blue tetrazolium/5-bromo-4-chloro-3'-indolylphosphate,p-toluidine salt (NBT/BCIP); the positive results were easily visualized as blue dots formatted on the microarray surface. The performance of a DNA microarray was tested against 14 representative collection strains and mock-contamination food samples. The combination of mPCR and a visual micro-plate chip specifically and sensitively detected Salmonella enterica, Shigella flexneri, E. coli 0157:H7 and Listeria monocytogenes in standard strains and food matrices with a sensitivity of similar to 10(2) CFU/mL of bacterial culture. Thus, the developed method is advantageous because of its high throughput, cost-effectiveness and ease of use.
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