Primers and kit for efficient triple detection of SVCV, IHNV and CEV

Abstract

The invention discloses a primer and a kit for efficient triple detection of SVCV, IHNV and CEV. An improved LAMP technology is used as a gene amplification reaction principle, gold nanoparticles are added into a reaction system, ssDNA and protease are adsorbed, non-specific reaction in the heating process is inhibited, the purpose of hot start is achieved, and the non-specific reaction in the heating process is avoided. An improved LAMP technology and a micro-fluidic chip technology are combined, the carp spring viraemia virus, the infectious hematopoietic necrosis virus and the carp edema virus can be rapidly and accurately detected and distinguished, meanwhile, a reaction reagent is pre-embedded in the micro-fluidic chip, same three-index detection is achieved, and user operation is easy 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 efficient triple detection of SVCV, IHNV and CEV. Background technique [0002] Spring viraemia of carp virus (SVCV for short), also known as carp rhabdovirus, belongs to the genus Vesivirus in the family Rhabdoviridae. There is a layer of envelope, the size of the virus is 180×70nm, and single-stranded RNA. Current studies have proved that carp spring viremia virus has only one serotype. The virus can mainly cause large-scale outbreaks of carp spring viremia in carps. [0003] Infectious haematopoietic necrosis virus (IHNV) belongs to Rhabdoviridae (Rhabdoviridae), Nora rhabdovirus (Novirhabdovirus), IHNV is the representative species of this genus. The IHNV virion contains a linear, antisense, single-stranded RNA of approximately 11 kb in length. Infectious hematopoietic necrosis disease primarily affects rainbow trout, including ste...

AI Technical Summary

Problems solved by technology

Using the existing PCR technology, because the reaction needs to be cycled in three 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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