Detection And Identification Of Food Borne Pathogens Using Oligonucleotide Microarray

The health of humankind is threatened by the pathogenic bacteria pollution of food which can lead to many serious diseases. The steps of conventional culture method and biochemical confirmation are trivial, and need many days to acquire the results. The technique of immunology and DNA probe hybridization also has the problems of specificity and sensitivity. PCR applied in detection of bacteriology has the characters of sensitivity, rapidity and specificity. However, the standardized PCR protocols can only detect a species of bacteria once time, and can't detect the samples polluted by many kinds of bacteria. In order to rapidly detect and identify the pathogenic bacteria in food, and simultaneously to provide scientific basis for clinical diagnosis, we developed the detection and identification system of pathogenic bacteria in food based on PCR and Oligonucleotide microarray. The primers chosen are located on the conserved region of 16S rRNA and 23S rRNA which is now used in bacterial taxonomy. The detected "targe" pathogens were Vibrio cholerae, Escherichia coli 0157:H7, Vibrio parahaemolyticus, Yersinia enterocolitica, Listeria monocytogenes, Shigella flexneri, Salmonella typhimurium, Campylobacter jejuni, Staphylococcus aureus. By using the universal primers labeled by fluorescein cy3/cy5, we can amplify the conserved stretches of DNA from any pathogenic bacteria in food. The oligonucleotide prodes designed in the variable region in 16S rRNA and 23S rRNA could be used to detect the pathogenic bacteria by by...