| Food safety was always concerned by governments and the people.It related the health and life of consumers.Food borne pathogens were the primary reasons leading to tbod poisoning.Thus,the detection for food borne pathogens was urgently needed.This research is based on common pathogenic pathogens in food,the molecular immunology,the molecular biology detection approaches were carried out.In the research,the immunity chromatographic analysis,PCR,the Real-time fluorescence PCR,the genetic chip were used for food borne pathogens detection.And these detection techniques were firstly applied to imported and exported commodity inspection.The newly developed detection protocols were for the first time in the world utilized to detect food borne pathogens.Which have momentous meanings for improving the imported and exported food inspection efficiency,reduce the time of inspection and remove the trade barriers which constituted by developed nations.The immunity chromatographic analysis technology was used for detecting staphylococcal enterotoxin A,B and C and Listeriolysin O.By using this technology, the inspection boxes having our own intellectual property rights were developed,and a set of rapid field inspection method was established.The method is specificity and more efficient than conventional methods.The establishment of this detection method provided a firm technical and theoretical basis.To detect food borne pathogens rapidly,the PCR technology was performed in this study.The results show that the PCR technology was availability for detecting food borne pathogens.The PCR method can greatly reduce the detection time,which have momentous meanings for cargo going through customs immediately.General food borne pathogens,such as the Salmonella,were known using the real-time fluorescence PCR approach to detect.By searching literature and using the isogenous sequence similarity BLAST analysis,the food borne pathogens target gene sequences were found.Based on these sequences,specific primers and probes for real time PCR detection were designed and selected.The systematically compared results between real time PCR method and GB,SN were identical.From these studies,the real time fluorescence PCR method is established.And the detection boxes were applied to imports and exports inspection,disease control and clinical diagnosis.The limit of detection was approximately 84cfu/mL -7000cfu/mL.In DNA extract experiment,the results of using boiled Salmonella enrichment broth and boiled single Salmonella bacterium as template of the real time PCR were as good as using Salmonella DNA. In the study of genetic chip of detection food borne pathogens,9 primers and 9 probes which are uniform in PCR detection profile and condition were selected and optimized,which could primers in one multiple PCR reaction to perform amplifying reaction simultaneously,so the interaction interferences between primer pairs in the process of multiple PCR reaction condition optimization,between every pairs of primers and products of multiple PCR reaction and between oligonucleotide length and structure of primers,probes and fluorescein labeled products were overcome.The studies showed 55℃was the best hybrid temperature for gene hybrid.Thus the protocols of high density food borne pathogens detection chip based on multiple PCR technique were established.The risk analysis of frozen aquatic products from Dandong port was carried out. The risk analysis mathematic model of food borne pathogens was established.The results showed that the risk probability of Listeria monocytogenes that take the frozen octopus,conch and crab,etc.for once were zero.The risk probability of Salmonella was 1.57×10-2,7.62×10-3,2.19×10-2 and 1.62×10-2 to health people.The Salmonella risk probability was 0.12,6.61×10-2,0.15 and 0.12 to susceptibility people. The risk probability of V.parahaemolyticus was 6.21×10-6,6.65×10-7,3.85×10-7,1.26×10-8,1.28×10-6 and 9.18×10-7.In this research,the food borne pathogens detection protocols based on molecular immunology,PCR,the real time PCR and gene chip were established for the first time in the world.And the risk analysis of frozen aquatic products from the North Korea was firstly carry out in the world.
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