Primers, kit and method for detecting Vibrio parahemolyticus by PSR (polymerase spiral reaction) isothermal amplification reaction

Abstract

The invention discloses primers, kit and method for detecting Vibrio parahemolyticus by PSR (polymerase spiral reaction) isothermal amplification reaction. A pair of detection primers Ft / Bt and a pairof acceleration primers IF / IB are designed according to specific target sequence tlh of Vibrio parahemolyticus, and their nucleotide sequences are shown as SEQ ID NO. 1 to 4; the primers ensure thatdetection results are reliable. The invention also provides a kit for detecting Vibrio parahemolyticus by PSR isothermal amplification reaction; the kit has high sensitivity, good specificity, good operational convenience, high operational speed, good result accuracy and reliability and low detection cost, and is suitable for application in small- to medium-sized units and field detection. The kitused herein to detect Listeria monocytogenes by polymerase spiral reaction; the method herein never causes time loss due to temperature changes, has low time consumption, the amplified product has noneed for gel electrophoresis, and after developing with a fluorescent dye, the results can be judged with eyes.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a primer, a kit and a method for detecting Vibrio parahaemolyticus by PSR isothermal amplification reaction. Background technique [0002] Vibrio parahaemolyticus is a halophilic bacterium that mainly exists in coastal seawater, seabed sediments, and seafood such as fish, shrimp, shellfish, and oysters. Many people cause food poisoning due to eating uncooked seafood contaminated by Vibrio parahaemolyticus, which has a high proportion and great harm in bacterial food poisoning. At present, most of the routine detection methods in our country have to go through the reaction process of selective culture, biochemical identification and serology. These experiments are cumbersome and take 5 to 6 days (d), and the detection rate is low, which brings difficulties to seafood production, entry-exit inspection and quarantine, and food hygiene supervision. There are reports of food poisoning c...

AI Technical Summary

Problems solved by technology

The gene chip detection and PCR amplification methods developed in recent years have strong specificity, rapidity and sensitivity, but the cost is high

Immunological detection methods, such as ELISA, immunochromatography, etc., have high sensitivity, but the operation process is complicated and the cost is high, so it is not suitable for large-scale detection of samples

At present, the most widely used constant temperature amplification technology-LAMP also has its limitations, such as complex primer design, high false positive rate, and high reagent price

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