Study on Detection of Five-kinds of Pathogenic Bacteria in Drinking Water by Multiplex-PCR / 微生物学通报

ABSTRACT

The five kinds of food-borne pathogenic bacteria including Salmonella sp.、Shigella sp.、Pseudomonas aeruginosa、Eterohaemorrhagic O157 and Vibrio rahaemolyticus that couldn't be detected in drinking water. The five pairs of primers were designed and composed according to the toxin genes, high-conservative genes and specific genes of these pathogens. The specificity of individual primer pairs in PCR was evaluated on DNA templates of more than thirty different bacterial isolates from ten species. Through optimizing the system and condition of multioplex-PCR, the detection sensitivity was improved dramatically. Multiplex-PCR analysis was primary performed on several water samples. The method established shortened the detection time and provided a cost-effective way in maximum. The experiment showed the multiplex-PCR using the five pairs of primers produced specific amplicons of expected sizes, the detection limits for the bacterial targets were estimated at 10~(1)~10~(2)cfu for one essay. It only took five to six hours to detect a sample, got the single, steady and clear result when the method was applied primarily in the analysis of water samples. The informative supplement to those areas which were environmental inspection, water detection, food sanitation supervision , commodity inspection and detection, etc.