Multiple PCR (Polymerase Chain Reaction) primer groups and detection method for synchronously detecting four pathogenic bacteria of siluriformes fish

Abstract

The invention discloses multiple PCR primer groups and a PCR detection method for synchronously detecting four pathogenic bacteria of a siluriformes fish. The multiple PCR primer groups and the PCR detection method have the advantages that high detection efficiency and accurate results are achieved, and the four pathogenic bacteria of the siluriformes fish can be synchronously detected. The multiple PCR primer groups comprise a catfish edwardsiella ictaluri primer pair, a flavobacterium cloumnare primer pair, an aeromonas hydrophila primer pair and an aeromonas punctata primer pair. The four pairs of PCR primer groups for different pathogenic bacteria are designed; one multiple PCR is established by utilizing the primer groups; the purpose of synchronously and quickly identifying the four pathogenic bacteria is realized; moreover, the sensibility is good, the specificity is high; relative to a conventional detection method, the cost needed by the PCR detection method is lower; the operation is simpler and more convenient; the needed time is shorter; the pertinence is high; the multiple PCR primer groups can be applied to the analysis and the detection of any one bacterial sample in multiple environments containing catfish edwardsiella ictaluri, aeromonas punctata, flavobacterium cloumnare and aeromonas hydrophila.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a primer sequence for simultaneously detecting four pathogenic bacteria of Siluriformes cultured fish, Edwardsiella silgi, Flavobacterium columnar, Aeromonas hydrophila and Aeromonas punctatus and detection methods. Background technique [0002] Siluriformes have become the focus of freshwater aquaculture development in the world: channel catfish are mainly raised in the United States; catfish, catfish and channel catfish are mainly raised in Europe; catfish and catfish are mainly raised in Africa; The southern largemouth catfish, leatherbeard catfish, bearded catfish, long-snout catfish, river yellow catfish, big fin catfish, etc. are being raised in large quantities as special aquatic products, and the current development momentum is very fast, among which yellow catfish is the most popular. Yellow catfish is a small fish that is widely distributed in major freshwater wate...

AI Technical Summary

Problems solved by technology

What's more troublesome is that DNA polymerases have different amplification efficiencies for different primers, and different amplified fragments in multiplex PCR will compete with each other for resources, often resulting in unbalanced amplification efficiencies, resulting in false negative results. High-efficiency fragments can be easily detected, while low-efficiency fragments are completely reduced to the background

At the same time, primer dimers are prone to appear, which brings difficulties to the detection

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