Primer and kit for efficient duplex detection of rabbit plague and rabbit plague type-2

Abstract

The invention discloses a primer and a kit for efficient duplex detection of rabbit plague and rabbit plague type-2, an improved LAMP technology is taken as a gene amplification reaction principle, gold nanoparticles are added into a reaction system to adsorb ssDNA and protease, a non-specific reaction in a heating process is inhibited, the purpose of hot start is achieved, and the non-specific reaction in the heating process is avoided; by combining the improved LAMP technology and a micro-fluidic chip technology, rabbit plague and rabbit plague type-2 can be rapidly and accurately detected and distinguished, and meanwhile, a reaction reagent is pre-buried in the micro-fluidic chip, so that the same double-index detection is realized, and the operation of a user is simple and convenient.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to a primer and a kit for high-efficiency dual detection of rabbit plague and rabbit plague type 2. Background technique [0002] Rabbit hemorrhagic disease (RHD), commonly known as rabbit plague, is an acute and highly fatal infectious disease of rabbits caused by rabbit hemorrhagic disease virus (RHDV). RHD is characterized by respiratory system hemorrhage, parenchymal organ congestion, swelling, hemorrhage and liver necrosis. It mainly harms many breeds of farmed and wild rabbits over 2 months old, with a mortality rate of 80%-100%. In 2010, a new species of RHDV, namely rabbit hemorrhagic disease virus 2 (RHDV2), appeared in France, which could infect and kill rabbits within 30 days of age and immunized rabbits with inactivated vaccines of traditional RHDV strains. The new rabbit viral hemorrhagic disease (newrabbit hemorrhagic disease, nRHD), also known as r...

AI Technical Summary

Problems solved by technology

Using the existing PCR technology, because the reaction needs to be cycled in two different temperature zones, the requirements for the instrument are high, and the cost is relatively high, and the PCR technology only has one pair of amplification primers, which is susceptible to interference and relatively insufficient in specificity, and It takes a long time to produce results, high professional requirements for operation, and many micro-dosing steps

The LAMP technology has 4 different specific primers, so the accuracy of the detection results is higher, but because it is a constant temperature reaction, it lacks a hot-start enzyme similar to PCR, and it is easy to produce non-specific amplification during the heating stage of the equipment, which will affect the detection results.

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