Development Of A Rapid Test Box For Common Food-Borne Pathogens

Bacterial food-borne diseases are still a major food safety issue in the world.The establishment of rapid food pathogen detection methods is essential to control the biological risks of agricultural products and food from production,transportation,storage,port customs clearance,sales and consumption.In recent years,rapid detection methods for bacteria in food have developed rapidly and have a wide variety of types.The biggest disadvantage of traditional methods is that they are time-consuming and cumbersome.They cannot detect contaminated samples quickly,and cannot find and control pollution sources in time,or detection sensitivity.Difficult to meet the requirements.In this research,a portable real-time fluorescent PCR instrument is combined with one-step rapid DNA extraction technology to form a portable rapid microbial detection container.Due to its inherent flexibility,the portable device is not limited to the laboratory,but can be performed anytime and anywhere,improving the detection efficiency.Timeliness and convenience provide good technical support for the application of small spaces,ports,remote areas,and on-site rapid detection,so that portable fluorescent quantitative PCR technology has more room for development.The standard strains of Cronobacterium,Salmonella,and Vibrio parahaemolyticus were used as research objects to establish a rapid detection system to achieve rapid detection of target strains.The specific research content was as follows:(1)Optimize the one-step rapid DNA extraction technology and study the rapid real-time fluorescence quantitative PCR instrument,respectively,using Cronobacterium,Salmonella,and Vibrio parahaemolyticus)the standard strain was the research object,and its DNA was extracted as a positive sample.The rapid detection system of the target strain was established.The traditional boiling method was used as the control,and the rapid real-time fluorescent PCR reaction program was established through optimization.The results showed: the establishment of a step the detection sensitivity of the method DNA extraction technology was equivalent to that of the traditional boiling method,and the established fast real-time fluorescent quantitative PCR method can shorten the traditional detection time from 82 min to about 30 min,and control the time required for the entire detection process to about 40 min.In order to meet the rapid and convenient detection requirements of on-site detection,the optimized rapid test system had the advantages of timeliness,convenience,quickness and simplicity,and high sensitivity.It has a good application prospect in the rapid detection of food-borne pathogens at ports.(2)The rapid detection method and the national standard method were used for the rapid detection method and the national standard method,and the comparison analysis found that: the rapid detection method the test results were consistent with the national standard method,and the rapid test method shortens the test time.Therefore,the rapid test system established in this study was suitable for the rapid detection of pathogenic bacteria in food and provides technical support for the detection of food-borne pathogens.(3)Integrate the rapid test system established in this study into a portable microbial rapid test box,and test the repeatability,stability and shelf life of reagents in the rapid test box.The research results showed that the rapid test box was repeatable and stable.All were good.The reagent can be stored for 6 months in the dark at-20°C,and the quick-check box was simple to operate,convenient to carry,and suitable for on-site detection of food-borne pathogens.