Detection method of prawn IHHNV (infectious hypodermal and hematopoietic necrosis virus) and used nucleic acid isothermal amplification detection kit

Abstract

The invention discloses a prawn IHHNV (infectious hypodermal and hematopoietic necrosis virus) nucleic acid isothermal amplification detection kit which comprises a grinding fluid tube containing a grinding fluid, a nucleic acid extracting solution tube A containing a sodium acetate solution, a nucleic acid extracting solution tube B containing absolute ethyl alcohol, a nucleic acid extracting solution tube C containing an alcohol solution with a mass concentration of 70 percent, a TE (tellurium) buffer solution tube containing a TE buffer solution, a UNG (uracil-DNA glycosidase) enzyme tube containing uracil DNA (deoxyribonucleic acid) glycosylase, an LAMP (Loop-mediated isothermal amplification) reaction liquid tube containing an LAMP reaction liquid, a BstDNA polymerase tube containing Bst DNA polymerase, a color-developing agent tube containing nucleic acid dye SYBR Green I, a positive control nucleic acid tube containing prawn IHHNV positive DNA and a negative control tube containing sterilizing double distilled water. The invention also provides a method for detecting the prawn IHHNV by utilizing the detection kit. The method has the characteristics of low cost, high detection sensitivity and easy site operation.

Description

technical field [0001] The invention relates to an aquatic animal pathogen detection technology, in particular to a nucleic acid isothermal amplification detection kit and a detection method for infectious subcutaneous and hematopoietic tissue necrosis virus of prawns. Background technique [0002] Infectious hypocutaneous and hematopoietic necrosis virus (IHHNV) was first discovered in the cultured Litopenaeus stylirostris in Hawaii. It is widely distributed and seriously harmful, causing a mortality rate of over 90%. It can also infect the cultured shrimps around the world. , is one of the other important diseases of crustaceans delineated by the International Office of Epizootics (OIE), and has a great impact on the development of shrimp farming in the world. Among the various viruses that can infect Penaeus vannamei, IHHNV can cause Runt-deformity syndrome (RDS for short). Although it will not cause a large number of prawns to die, the virus infection will actually cause...

AI Technical Summary

Problems solved by technology

Although electron microscopy can directly observe the presence of virus particles, the operation is complicated, time-consuming and costly

The HE staining method is based on pathological techniques, which cannot directly detect the virus, but can only be detected by using the histopathological signs of the disease

Antibody detection method and nucleic acid probe hybridization method are used to detect the protein or nucleic acid components of the virus itself. The detection sensitivity is low and time-consuming, and can only be used for the detection of diseased shrimp or shrimps that are about to become diseased; IHHNV has not yet caused infection Or it is difficult to detect with the above methods in the very early stage of infection

Although the PCR detection method of IHHNV overcomes the shortcomings of the first four methods, it can achieve relatively fast and accurate detection of IHHNV under laboratory conditions, but because conventional PCR detection requires expensive PCR machines, gel electrophoresis and imaging systems , and the detection time is slightly longer, which also greatly limits the popularization and application of PCR detection method in production practice