Multiplex-polymerase chain reaction (PCR) detection method capable of simultaneously detecting vibrio parahaemolyticus, vibrio vulnificus and vibrio alginolyticus

Abstract

The invention discloses a multiplex-polymerase chain reaction (PCR) detection method capable of simultaneously detecting vibrio parahaemolyticus, vibrio vulnificus and vibrio alginolyticus. The detection method comprises the steps of firstly, preparing a deoxyribonucleic acid (DNA) template of a detected object to put into a reaction tube; putting three vibrio primers into the reaction tube in step a to carry out multiplex-PCR reaction, wherein the primer concentrations are 1-20pmol / mu l, 1-20pmol / mu l and 1-30pmol / mu l in sequence; the primer sequences of the vibrio parahaemolyticus are 5'-GCAGCTGATCAAAACGTTGAGT-3', and 5'-ATTATCGATCGTGCCACTCAC-3'; the primer sequences of the vibrio vulnificus are 5'-TTCCAACTTCAAACCGAACTATGAC-3', and 5'-ATTCCAGTCGATGCGAATACCGAATACGTTG-3'; and the primer sequences of the vibrio alginolyticus are 5'-CGAGTACAGTCACTTGAAAGCC-3', and 5'-CACAACAGAACTCGCGTTACC-3'.

Description

technical field [0001] The invention relates to a multiplex PCR detection method, in particular to a multiplex PCR detection method which is short in time consumption, high in efficiency and high in sensitivity and can be directly applied to clinics and can simultaneously detect Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio alginolyticus method. Background technique [0002] Vibrio is widely distributed in nearshore and estuary sea areas, and most of them are dominant flora that exist on the surface and intestinal tract of seawater and marine organisms, and are also one of the most important pathogenic bacteria that cause bacterial diseases in marine animals. A large amount of research and production practices have shown that vibriosis is a type of bacterial disease caused by Vibrio bacteria, which is widely prevalent and has a high incidence rate. The main pathogenic Vibrio species are: Vibrio alginolyticus ), Vibrio parahaemolyticus ( Vibrio parahaemolyticus ), ...

AI Technical Summary

Problems solved by technology

[0006] At present, multiplex PCR is mainly used in the detection of Vibrio that can cause human diseases, such as food testing, etc., but in aquatic animal vibriosis, the common PCR method is basically used, and only one type of Vibrio can be detected at a time, and the time required for detection Long and often results in missed or false detections

[0007] The present invention aims to solve the problem that the PCR method in the prior art can only detect one type of Vibrio at a time, which takes a long time to detect and often causes missed detection, and provides a A multiplex PCR method that can simultaneously detect multiple vibriosis pathogenic bacteria

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