Method for detecting pathogenic bacteria in foods by isothermal rapid amplification

Abstract

The invention discloses a method for detecting pathogenic bacteria in foods by isothermal rapid amplification and belongs to the technical field of food safety detection. The method comprises the following steps: determining detection target nucleic acid aiming at each bacterium through multiple sequences; designing a specific molecular beacon probe and a primer; taking a genome DNA (Deoxyribonucleic Acid) of each strain as a template and establishing real-time fluorescence CSDPR; carrying out the isothermal nucleic acid amplification and identifying the specificity, sensitivity and minimum limit of detection of a CSDPR method. The detection of an actual sample determines that the method established by the invention has practical feasibility. The method disclosed by the invention has commonality on detection of various pathogenic bacteria in the foods and the pathogenic bacteria comprise staphylococcus aureus, salmonella, enterohemorrhagic escherichia coli, shigella and the like; only if a specific part in the molecular beacon probe needs to be replaced, effective detection of the different pathogenic bacteria can be realized; and the method has high detection sensitivity and high specificity, simple principle and operation and the pretreatment of the sample is simple.

Description

technical field [0001] The invention relates to a method for isothermal rapid amplification detection of pathogenic bacteria in food, belonging to the technical field of food safety detection. Background technique [0002] In recent years, in the context of globalization, with the rapid growth of global food trade, the global flow of population, and the emergence of drug-resistant strains, the incidence and fatality rate of food-borne diseases have gradually increased, and have become the world's food Important issues in the fields of safety and public health. Microorganisms (including bacteria, viruses, fungi, and parasites) that spread through contaminated food or water and cause foodborne illness are called foodborne pathogenic microorganisms. In foreign countries, in the cases of foodborne diseases caused by pathogenic microorganisms, the main pathogenic factor is pathogenic bacteria (or foodborne pathogens), and the most common foodborne pathogens include enterohemorrh...

AI Technical Summary

Problems solved by technology

The relevant PCR method detects Staphylococcus aureus at the molecular level, which has high specificity and sensitivity, especially the real-time fluorescent quantitative PCR method, which can detect the reaction process of the system in real time and avoid the contamination of carcinogens such as EB during the experiment , so it has certain advantages in the detection method, but the reaction process requires expensive and complicated instruments, the detection cost is high, it is difficult to achieve basic level, and the scope of application is limited

[0005] In addition, there is currently a lack of methods that can simultaneously and rapidly quantitatively detect multiple food-borne pathogens such as Staphylococcus aureus, Salmonella, enterohemorrhagic Escherichia coli, Shigella, etc.

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