Quadruple real-time fluorescence PCR method for detecting four types of pathogenic bacteria

Abstract

The invention discloses a quadruple fluorescence PCR method for detecting four types of pathogenic bacteria. The four types of pathogenic bacteria are staphylococcus aureus, salmonella enterica, listeria monocytogenes and vibrio parahemolyticus. Four elements, namely sample DNA, a pair of universal primers and four probes, fluorescence PCR premix liquid and positive control are adopted in the method. The method comprises the following steps: designing the primers and the probes in accordance with 16S rRNA genes of the four types of bacteria, implementing amplification on a target sequence and exciting corresponding fluorescence signals; and preparing a kit from the to-be-detected sample DNA in accordance with a reagent formula and amplification conditions of the fluorescence PCR amplification; and specifically, the method comprises the following steps: establishing a quadruple fluorescence PCR primer system for detecting the four types of bacteria; establishing the quadruple fluorescence PCR kit for detecting the four types of bacteria; and determining the quadruple fluorescence PCR identification method for detecting the four types of bacteria. The method provided by the invention has the advantages of being low in standard error, short in detection time, and being capable of simultaneously detecting a plurality of target genes and reducing reagent cost; and the method is applicable to detection of pathogenic bacteria in food and cosmetics.

Description

technical field [0001] The invention relates to an identification method including genes, a detection method of pathogenic bacteria in food and cosmetics, in particular to a detection method of pathogenic bacteria in instant food. Background technique [0002] People depend on food, and food safety is one of the basic pillars of social harmony. In terms of food and cosmetics, especially in the field of ready-to-eat food, the detection of pathogenic bacteria is one of the important indicators of food safety. Referring to decades of accumulated experience in disease control and prevention, currently in various product implementation standards and food and drug supervision and sampling rules at all levels, most of the pathogenic bacteria detection items in ready-to-eat foods are Staphylococcus aureus and Salmonella; The main component is the detection of increased Listeria monocytogenes in livestock and poultry products; the main component is the detection of increased Vibrio ...

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Problems solved by technology

These patents are similar to academic articles, with a wide variety of reagents, complex operation steps, and high requirements for the skills of experimenters, and are more suitable for scientific research; The ability of personnel is uneven, and higher requirements are put forward for reagent cost, operation simplicity, method standardization, etc.

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