Establishment Of The Detection Technology System Of Pathogenic Bacteria In Heat-processed Food And Aquatic Products

The kinds of pathogenic bacterium in heat-processed food and aquatic products are the fatal factors to threat the food safety and physical health of human being.Currently,the domestic and foreign inspection method to the heat-processed food and aquatic products mostly be in the lever of low efficiency and single target.This research has materialized the system study on the high throughput technology like the high throughput inspection,toxin code gene filter,quick classifying,pathogenic bacterium mutant cluster inspection,and identification diagnosis of kinds of pathogenic bacterium of heat-processed food and aquatic products through PCR combing high throughput nuclear acid inspection method of denaturing high performance liquid chromatography,DHPLC and get the following test results by building the corresponding technology platform.1.The high throughput technology of three kinds of foodborne pathogenic bacterium like the common bacillus cereus,Clostridium bobulinm and cl.perfringens and its toxin code gene which aim at the heat-processed food has been developed with the application of PCR combining with the principal of separating DNA fragment under non-denaturing condition by denaturing high performance liquid chromatography,DHPLC and through the optimization of multi-PCR and DHPLC analysis conditions.As well as the specific gene inspection platform to detect the pathogenic bacterium of heat-processed food through PCR-DHPLC inspection has been established.The researched and developed multi PCR-DHPLC to inspect the three kinds of sporogenes under non-denaturing condition is creative.2.The high throughput inspection technical difficult of common Lactobacillus PCR-DHPLC in heat-processed food has been solved and the Lactobacillus quick inspection technology has been developed.With the application of PCR-DHPLC non denaturing analysis condition,based on the inspection level of genus,the food lactobillus inspection method has been developed,which remedies the problems of error and leak inspection caused by long period,unstable reagent quality and biochemical reaction of common culture identifying,and provides a scientific,high efficient and quick inspection medium to the lactobillus quick inspection during the food safety and industry production inspection.3.The high throughput strain-classifying technical difficult to reach classification identification between lactobillus strain relying on the difference of general primer and DNA nucleotide base, the high throughput strain-classifying identifying method of the nine classified strain of lactobillus has been developed with the application of high throughput type-classifying identifying method under DHPLC partial denaturing condition and aiming at the nine classified strains of lactobillus,the scientific,high efficient and quick identifying purpose of the type-classifying of nine classified strains of lactobillus can be identified at one time by one enrichment,one inspection has been really reached.The type-classifying identification technology of nine strains of lactobillus by DHPLC inspection under partial denaturing condition is creative. 4.The technical difficult of high throughput quick inspection to the common pathogenic bacterium of aquatic products has been solved and the high throughput quick inspection technology of completing the inspection from kinds of target bacterium,one time compound enrichment and one time PCR-DHPLC has been created.Taking the nine microorganisms like the common pathogenic bacterium,comma bacillus,V.parahemolyticus,V.vulnificus,V. alginnolyficus,V.mimicus,enterohemorrhagic escherichia coli 0157:H7,salmonella,Listeria monocytogenes and staphyloccocus aureus rosenbach in aquatic products as research target, this reach has developed the PCR-DHPLC quick inspection under non-denaturing condition and the specific gene inspection platform to detect the pathogenic bacterium of heat-processed food through PCR-DHPLC inspection,and carried out the filter inspection to of comma bacillus mutant cluster.Nine kinds of pathogenic bacterium of aquatic products are inspected together at one operation process has made the inspection time,quantity and level greatly leaped.Therefore to reach the quick,accurate and high sensitive inspection purpose to the abovementioned nine kinds of microorganism.The researched and developed multi PCR-DHPLC to inspect the nine kinds of common pathogenic bacterium of aquatic products under non-denaturing condition is creative.Above all,this research has firstly developed the DHPLC accurate and high throughput inspection technology of kinds of pathogenic bacterium of heat-processed food and aquatic products,quick type-classifying identifying technology of pathogenic bacterium,toxin code gene filter technology as well as filled in the vacancy of this research.Meanwhile,it has standardized the inspection technology,and formulated the sector standard of Entry-exit inspection and quarantine of P.R.C based on the research basis;and the research achievement has been transformed and declared the state invention patent with important research meaning and application value.