Specific target, primer, detection method and application for detecting vibrio parahaemolyticus

Abstract

The invention provides a specific target for detecting vibrio parahaemolyticus, a primer pair, a detection method and application. According to the method, specific gene sequences capable of being used for detection in vibrio parahaemolyticus are mined according to a primary screening result of a local database and a secondary screening result of online comparison, the mined specific target gene is combined with a traditional target gene to construct a dual real-time fluorescence PCR method, and PCR primer pairs are designed according to a self-screening target gene VPA1585 and a traditional target gene tlh. In order to ensure high specificity and sensitivity of the detection method, a dual real-time fluorescent PCR method is adopted, a PCR reaction system is reasonably optimized, and a third-generation saturated fluorescent dye SYTO Green 9 is innovatively used for replacing an unsaturated fluorescent dye SYBR Green in a traditional real-time fluorescent PCR system, so the vibrio parahaemolyticus can be rapidly and accurately identified.

Description

technical field [0001] The invention belongs to the technical field of food safety detection, and specifically relates to a specific target for detecting Vibrio parahaemolyticus, primers and applications based on the specific target VPA1585 and the traditional target tlh, and the combination of VPA1585 and tlh to construct The dual real-time fluorescent PCR system is a method for detecting Vibrio parahaemolyticus. Background technique [0002] Vibrio parahaemolyticus (Vibrio parahaemolyticus) is a human pathogen that is widely distributed in the marine environment and is often isolated from a variety of raw seafood. Consumption of unprocessed or undercooked seafood products contaminated with Vibrio parahaemolyticus May cause food poisoning and lead to the development of acute gastroenteritis, although gastroenteritis caused by Vibrio parahaemolyticus is usually self-limited, it can be dangerous in people with underlying medical conditions such as liver disease or immune dise...

AI Technical Summary

Problems solved by technology

At present, the commonly used detection targets of Vibrio parahaemolyticus are tdh, trh, tlh, groEL and toxR genes, but the above-mentioned genes are found to have certain limitations or false detection rates in the actual application detection process, which makes the detection efficiency and accuracy The degree is reduced, and the advantages of the PCR method cannot be fully utilized

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