The Sensitivity Test And Preservation Test Of 5 Pathogenic Bacterium Detection Kit Chips In Pork And Related Products

Biochip is microarray of high-density organize,cell,protein,nucleinic acid,carbohydrate and other biont constituent covered on slice of silicon,nylon membrane and other solid phase planks.Making chip be hybridization with tagged sample,analyze the biological sample by detecting the hybridization signal.Biochip technique as the life science core technique in 21 century is not only using for identification of bacteria,but also using for the express of detecting gene,researching of biosystem reaction during versity,researching of biosystem reaction for light,enviroment detecting,discover new gene and other fields with bright perspective.Especial the wide and wide using for microorganism assay in animal products.This article is about detection kit chip conservation test and sensitiveness test for salmonella,Listeria monocytoge,Staphylococcus aureu,C.jejuni,Bacillus coli O157:H7 in import and export pork and related products,which provide science evidence for the application condition amd expiration date ofbiochip during fast detection of this 5 varietas pathogenic bacteria in import and export pork and related products.According the disign of conservation,this experiment put microbiological detection kit chip on 37℃enviroment,using the 5 varietas pathogenic bacteria(almonella,Listeria,Staphylococcus aureu,C.jejuni,Bacillus coli O157:H7) which can be detected by kit to detect the chip stability.From the first day,take chip saving on 37℃enviroment and put 4℃enviroment saving with evacuation,in 7 day take all chips on 4℃enviroment outaccording the indication,take the 5 varietas pathogenic bacteria to do genome DNA extraction,PCR amplification,PCR products GE separately.At the same time making the mixture of 5 bacteriaes PCR products hybridization with saving chip in 2 hours under 42℃temperature.After wash chip,using PFMicrobeⅡSystem get the result.Experiment take 7 days which is equal chip saving 10 month and 15 days under 20℃enviroment.The result shows that significant difference between these 5 bacteriaes test date and detectability,and the signal intensity is stronger than detectability,results all are masculine.The approve that microbiological detection kit chip can be saved in 7 days under 37℃,that means save 10 month and 15 days under 4℃enviroment,they can keep stability and activity.According the sensitiveness test design,using artificial culture bacteriaes do detection test,take Staphylococcus aureus,C.jejuni,Bacillus coliO157:H7,almoneUa,Listeria 5 bacteriaes.Inoculating collective media nutrient broth,cultivate 16h overnight,take the bacteriae fluid do concentration measurement in the next day,from quantitation bacteria fluid extract genomic DNA,and doing quantitate test,extract 5ng,2.5ng,lng,0.5ng separately to do PCR amplification,electrophoresis detecting PCR amplification products,to which do hybridization detection.The result shows the kits' sensitiveness have slender difference between different bacteriaes,but all are above 8.7×10~4 cfu,Staphylococcus aureus,C.jejuni,Bacillus coliO157:H7,almonella,Listeria and C.jejuni all can detect the target bacteriaes,but the sensitiveness of Bacillus coliO157:H7 and Listeria are higher.So the result is the kit sensitiveness is 8.7×104 cfu/mL after doing enrichment culture for samples.