Primers and reagent kit for rapidly detecting specific vibrio parahemolyticus virulence plasmid pVA1 on site

Abstract

The invention discloses primers and a reagent kit for rapidly detecting the specific vibrio parahemolyticus virulence plasmid pVA1 on site. The sequences of the primers are shown as SE ID NO:3-6. By the adoption of the primers, the specific vibrio parahemolyticus virulence plasmid pVA1 can be easily, conveniently, rapidly and visually detected, accuracy is high, specificity is good, detection time is short, and operation is easy and convenient.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer and a kit for on-site rapid detection of specific Vibrio parahaemolyticus virulence plasmid pVA1. Background technique [0002] Vibrio parahemolyticus (Vibrioparahemolyticus) is a Gram-negative bacillus with arc-shaped, rod-shaped, filamentous and other shapes without spores. At present, the PCR methods used to detect the bacteria mainly include arbitrary primer PCR (arbitrarilyprimedPCR, Ap-PCR), conventional PCR, multiplex PCR (multiplex-PCR), real-time PCR (Real-timePCR). [0003] The thermolabile hemolytic toxin tlh gene was discovered by Hatsumi et al. in 1985. Both clinical isolates and environmental isolates have tlh gene, which is species-specific. Therefore, its coding gene tlh gene is often used as the target gene for detection of Vibrio parahaemolyticus. However not all V. parahaemolyticus are equally pathogenic. Thermostable direct hemolytic toxin TDH and ther...

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Problems solved by technology

[0006] At present, the most accurate means of detecting this specific Vibrio parahaemolyticus is the method of PCR detection of pVA1 plasmid established in 2013. This conventional PCR detection method requires a series of PCR instruments, electrophoresis instruments, ultraviolet detectors, computers, etc. Professional detection equipment; at the same time, the detection process needs to draw products from the reaction system for electrophoresis or Southern hybridization tests for identification. During the transfer process, it is easy to contaminate, causing false positives, and time-consuming. It is suitable for detection and research by specialized testing institutions. It is not conducive to the rapid monitoring and detection application of the farmers on the shrimp farm site

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