Prawn white spot syndrome virus nucleic acid isothermal amplification detection reagent kit and detecting method

Abstract

The invention relates to a detection kit for isothermal amplification of nucleic acid for the white spot syndrome virus and a detecting method thereof. The kit is designed by using a set of LAMP primers as the main body designed according to the gene conserved sequence of the white spot syndrome virus. The kit is provided with eleven agents needed by WSSV detection including SEMP lapping liquid, nucleic acid extract, UNG enzyme, TE buffer solution, enzymolysis buffer solution, LAMP reaction solution and on the like, thus realizing a programmed and standardized detection; therefore, the invention has the advantages of the highest sensitivity, simplicity, rapidity, safety, good specificity and low cost, and the amount of the copies detected with virus can be as low as 26. Only one water-bath is needed to correctly detect the white spot syndrome virus in prawns and aquaculture water within 2 hours by the kit and the detecting method. Moreover, the problem that the detection is subject to interference in the LAMP technology is also solved. The detecting method of the invention is expected to substitute the previous related detecting methods of the white spot syndrome virus, such as the electron microscope method, the TE staining method, the biopsy method, the antibody detecting method, the nucleic acid probe hybridization method and the PCR detecting method.

Description

technical field [0001] The invention relates to a marine biological pathogen detection technology, in particular to a nucleic acid isothermal amplification detection kit and a detection method for white spot syndrome virus (WSSV, White Spot SyndromeVirus) of prawns. Background technique [0002] Shrimp white spot syndrome virus is a large baculovirus that can widely infect seedlings, adults and broodstock of decapod crustaceans such as shrimp and crab. A large number of deaths have caused huge economic losses to the aquaculture industry. Therefore, it is of great significance to develop new technologies to detect WSSV quickly, accurately and sensitively, to strengthen the quarantine of seedlings and adult shrimp, and to monitor the environment of aquaculture water bodies, to prevent virus infection, cut off virus transmission, and ensure the healthy and sustainable development of my country's shrimp aquaculture industry. [0003] At present, there are mainly six detection m...

AI Technical Summary

Problems solved by technology

Although the existence of virus particles can be directly observed by electron microscopy, the operation is complex, time-consuming, and inaccurate; both TE staining and pathological sectioning are based on pathological techniques, which cannot directly detect viruses and can only be detected by using The histopathological signs of the disease are detected; the antibody detection method and the nucleic acid probe hybridization method are used to detect the protein or nucleic acid components of the virus itself. For detection, WSSV has not yet caused infection or is difficult to detect with these two methods in the very early stage of infection; although the PCR detection method of WSSV overcomes the shortcomings of the first five methods, it can achieve relatively rapid detection of WSSV under laboratory conditions. , accurate detection, but because conventional PCR detection requires expensive PCR machines, gel electrophoresis and imaging systems, this greatly limits the popularization and application of PCR detection methods in production