Study On Rapid Detection Of Eight Kinds Of Food-borne Pathogen By ATP Bioluminescence Method

The quantity control of food borne pathogenic bacteria was an important step in food testing, and it was also the key step to affect food quality and consumer safety. In recent years, ATP fluorescence detection technology had been applied in the field of food, medical and health, environmental assessment and so on. This paper was based on the egith kinds of common foodborne pathogenic bacteria, and mainly studied the applicability of the ATP fluorescence technique to detect food-borne pathogenic bacteria, the influence of differences of pathogenic bacteria ATP content on fluorescence technology and the practical of technology of continuous real-time detecting microbial content.At the same time, based on the principle of ATP luminescence,this paper puts forward the design scheme of ATP fluorescent pen,combined with two kinds of selective medium to achieve the purpose of preliminaryqualitative detection of pathogenic bacteria. Specific research contents are as follows:(1) In this paper, basing on ATP bio luminescence and supplementing by plate count method, we constructed the mathematical model of egith kinds of common foodborne pathogens bacteria concentration with the corresponding luminescence intensity.We compared the difference of ATP content between different strains, and the reliability of the bacteria count bioluminescence evaluation. The results showed that the linear correlation coefficient of egith kinds of food borne pathogens working curve,which in addition to enteropathogenic Escherichia coli was 0.8612, the rest were greater than 0.93. For the same fluorescence values , the different species of pathogenic bacteria contained in the total number of colonies were not significantly different. Comprehensive experimental data: ATP bioluminescence technology could quantitatively predict the number of egith kinds of food borne pathogens, which provides a theoretical support for the application of ATP fluorescence technology in food quality and safety.(2) We selected three kinds of pathogenic bacteria as the research object, basing on the principle of ATP bioluminescence,the standard curve about colony concentration with fluorescence intensity of food borne pathogenic bacteria were constructed in different growth stages.We calculated the four growth stages of ATP content. The results showed that the ATP content of three kind of food borne pathogenic bacteria in different growth stages had significant influence on colony counting.The standard curve could not be built by single growth period as a response to specific testing standards, and samples were repeatedly measured to establish mathematic model.(3) A selective enrichment broth (SSVP) was formulated to allow simultaneous growth of Salmonella spp., Proteus vulgaris, Staphylococcus aureus, and Vibrio parahaemolyticus in order to achieve rapid enrichment of these four food-borne pathogenic bacteria in aquatic products. At the same time, basing on SSVP, we added specific bacteriostatic agent and controlled bacteria time to achieve the goal of distinguishing the four kinds of food-borne pathogens. It showed as flows : (1) SSVP had a different degree of inhibition to non-target bacteria. When the initial inoculation concentration was 1CFU/mL, four kinds of target bacteria in single bacteria and mixed bacteria could reach 104CFU/mL after cultivating 8h? 12h respectively. It meet the modern testing technology. (2) We controlled enrichment time as 16h, 14h, when we could specifically detect of salmonella enteritidis and staphylococcus aureus in SSVP- sodium thiosulfate and SSVP- nalidixic acid respectively. During the time,high background bacteria was in demurrage, and low concentration target bacteria could achieve the most in the bacteria. SSVP could rapidly enrichment of four kinds of foodbome pathogenic bacteria , and two selective medium could qualitative detection of foodborne pathogenic bacteria. There was a ATP fluorescence detection pen system design, combined with the above medium, and it was expected to design a suitable for civilian and commercial ATP fluorescent detection pen.As a simple and effective method, the ATP fluorescence detection technology could be used to estimate the quality and safety of food quickly and accurately.