PCR specific amplified primer for fast detection of vibrio parahaemolyticus based on SCAR molecular markers and detection kit

Abstract

The invention discloses a PCR specific amplified primer for the fast detection of vibrio parahaemolyticus based on SCAR molecular markers and a detection kit. The nucleotide sequences of a forward primer and a reverse primer are as follows: the nucleotide sequence of the forward primer VpaF is 5'-ACTACCGAACACGGGCTGTAT-3'; and the nucleotide sequence of the reverse primer VpaR is 5'-GGCTGGGCATCGTAATCAT-3'.The kit comprises a mixed liquid of the PCR specific amplified forward primer VpaF and reverse primer VpaR dissolved in sterile double distilled water, wherein the concentration is 0.5-2 mu M. Cultured vibrio colony can be directly used to be subject to the PCR reaction for detection without extracting DNAs of the vibrio genome, thereby achieving the effects of time saving, labor saving, convenience, high speed, accuracy, good specificity and high sensitivity. The kit can be used for tracing the detection of the vibrio parahaemolyticus in the marine animal culturing process and monitoring the briny environment, thereby achieving high use value.

Description

Technical field: [0001] The invention relates to a PCR-specific amplification primer and a detection kit for detecting pathogenic bacteria in seawater cultured animals, in particular to a SCAR (Sequence characterized amplified region, characteristic sequence detection kit) with high detection accuracy and sensitivity, short time and low cost. Amplification region) PCR-specific amplification primers and detection kits for rapid detection of Vibrio parahaemolyticus by molecular markers. Background technique: [0002] Vibrio is the main conditional pathogenic bacteria of marine animals. Pathogenic Vibrio can cause explosive death of farmed animals. Together with viruses, other bacteria or parasites, it will cause huge damage to marine animals. It is recognized as It is one of the most serious diseases in mariculture. For example, Vibrio parahaemolyticus can cause large-scale deaths of farmed animals such as fish, shrimp, shellfish, and sea cucumbers, and it also brings hidden ...

AI Technical Summary

Problems solved by technology

For example, Vibrio parahaemolyticus (Vibrioparahaemolyticus) can cause large-scale deaths of farmed animals such as fish, shrimp, shellfish, and sea cucumbers, and it also brings hidden dangers to human health who accidentally eat food infected with Vibrio parahaemolyticus

At present, the detection methods for Vibrio include physiological and biochemical identification technology, enzyme-linked immunosorbent assay, 16S rRNA gene identification, etc., respectively have the following shortcomings: physiological and biochemical identification technology is time-consuming and laborious, and the detection accuracy is low; enzyme-linked immunosorbent assay The sensitivity is not high; 16S rRNA gene identification can only identify the pathogenic Vibrio to the genus, but not to the species

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