Efficient quintuplet primer for detecting shrimp diseases and kit

Abstract

The invention discloses an efficient quintuplet primer for detecting shrimp diseases and a kit. According to the primer and the kit, an improved LAMP technology serves as a gene amplification reaction principle, and gold nanoparticles are added into a reaction system to adsorb ssDNA and protease and inhibit nonspecific reactions during heating, thus, the aim of warm start is achieved, and nonspecific reactions during heating-up are avoided; and by combining the improved LAMP technology and a microfluidic chip technology, an accurate detection result can be presented rapidly, and the aim of joint detection on the same sample with a plurality of different indexes is achieved; meanwhile, reaction reagents are pre-buried into a microfluidic chip, five-index detection on one same is achieved, a user only needs to add a sample, and operations are simple and convenient; and the instrument is equipped with lithium batteries, so that on-site rapid detection is facilitated.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to a highly efficient five-way detection method for shrimp diseases (prawn infectious subcutaneous and hematopoietic necrosis virus (infectious type), prawn infectious subcutaneous and hematopoietic tissue necrosis virus (general type), yellow head virus , Stealing wild Tamura virus, Taura virus) primers and kits. Background technique [0002] Infectious hypodermal and hematopoietic necrosis virus (IHHNV), also known as Litopenaeus stylirostris densovirus and decapod densovirus, is the smallest virus among known prawn viruses, with a particle size of 22nm. Non-encapsulated icosahedron, linear single-stranded DNA, about 3.9kb in length, containing 3 open reading frames. IHHNV genotypes are divided into infectious and non-infectious types, of which infectious types include types 1 and 2; non-infectious types include types 3A and 3B. IHHNV is widely prevalent in th...

AI Technical Summary

Problems solved by technology

PCR technology is more classic, but its reaction needs to be cycled in two different temperature zones, which requires high equipment and relatively high cost. At the same time, PCR technology only has one pair of amplification primers, which is susceptible to interference, relatively insufficient specificity, and The result time is long, the operation professional requirements are high, and there are many steps for adding micro-dosage

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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