Fish pike fry rhabdovirus nested PCR detection primer group, kit and method and application

Abstract

The invention relates to a fish pike fry rhabdovirus nested PCR detection primer group, kit and method and application, and belongs to the technical field of molecular biology. The fish pike fry rhabdovirus nested PCR detection method comprises the steps that total RNA of a sample to be detected is used as a template, a reverse transcription reaction is carried out, and cDNA of the sample to be detected is obtained; the cDNA of the sample to be detected is used as a template to perform a first round PCR amplified reaction; then a first round PCR amplified product is used as a template to perform a second round PCR amplified reaction; and a second round PCR amplified product is subjected to agarose gel electrophoresis, gel is placed into an imaging system for observing whether a positive amplification stripe appears or not, and whether the sample to be detected contains fish pike fry rhabdovirus or not is judged. The fish pike fry rhabdovirus nested PCR detection method has the characteristics of high sensitivity and high specificity, and fish pike fry rhabdovirus-snakehead vesicular stomatitis virus can be rapidly and accurately detected.

Description

technical field [0001] The invention relates to a primer set, a kit, a detection method and an application for fish rhabdovirus nested PCR detection, and belongs to the technical field of molecular biology. Background technique [0002] PCR stands for Polymerase Chain Reaction technique, a process similar to DNA replication in reference cells. This technology uses DNA as a template, adding PCR enzymes and corresponding specific primers to perform programmed reactions in a PCR instrument, and copy and amplify to generate new complementary DNA fragments. The process consists of three steps: denaturation-annealing-extension. After multiple cycles, DNA amplification can be amplified millions of times within 2-3 hours. Because of its advantages of high amplification efficiency, strong specificity and high sensitivity, it has been widely used in the field of molecular biology research. [0003] Nested PCR is an improved solution based on the common PCR method. The difference fro...

AI Technical Summary

Problems solved by technology

In recent years, the virus has caused great harm to some aquaculture areas. In some artificially cultured ponds, the diseased fish had no obvious symptoms at the initial stage of infection, and their food intake was weakened and their physical fitness declined. Protruding, floating on the edge of the pool, sometimes rolling over, and soon died, there is no effective treatment

There is currently no special literature and technology for the identification of this virus, and ordinary PCR is generally used for detection, which has low specificity and sensitivity and a high probability of false positives

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