Real-time fluorescent quantitative PCR (polymerase chain reaction) detection kit for shrimp white spot syndrome virus

Abstract

The invention discloses a real-time fluorescent quantitative PCR (polymerase chain reaction) detection kit for shrimp white spot syndrome virus. The kit comprises the following reagents of a reagent A, a reagent B, a reagent C, a reagent D, a reagent E, a reagent F, a reagent G, sterilized double distilled water, and a DNA (deoxyribonucleic acid) absorption column, wherein the reagent A is prepared by adding water into hexadecyl trimethyl ammonium bromide, NaCl (sodium chloride), ethylene diamine tetraacetate, and hydroxymethyl aminomethane-hydrochloric acid, the reagent B adopts protease K, a solvent of the reagent B is the sterilized double distilled water, the reagent C is an ethanol water solution, the reagent D is prepared by mixing a PCR reaction buffer containing MgCl2 (magnesium chloride), Taq (thermos aquaticus) DNA polymerase and the sterilized double distilled water, the reagent E is prepared by mixing a primer P1, a primer P2 and deoxyribonucleotide, the reagent F is a fluorescent probe premixed solution, and the reagent G adopts a shrimp white spot syndrome virus positive plasmid. The kit has the advantages that the detection sensitivity is high, the kit is used for the early infection and tracking monitoring of the cultivation shrimp white spot syndrome virus, and the kit is also used for the related basic scientific research of virus multiplication.

Description

technical field [0001] The invention relates to a detection kit and a detection method of prawn white spot syndrome virus. Background technique [0002] Since the 1990s, shrimp white spot syndrome virus has caused acute and fatal infectious diseases of shrimp farming worldwide, causing huge economic losses to the shrimp farming industry. Shrimp white spot syndrome virus is an enveloped double-stranded DNA virus. In addition to prawns, the virus also infects marine and freshwater crustaceans such as crabs and crayfish. At present, the shrimp disease caused by the virus has become a worldwide epidemic disease. It is not only a huge threat to the shrimp farming industry, but also endangers the entire marine ecological environment. There is currently no effective treatment for crustacean viral diseases, and early detection and diagnosis of viruses is still the most important means of prevention and control. Therefore, the establishment of rapid, sensitive and practical detect...

AI Technical Summary

Problems solved by technology

Since the dye can bind to all DNA double strands and has no selectivity for the template, the specificity is poor and the quantification is relatively inaccurate

Wang Zhongfa and others initially established the Taqman real-time fluorescent quantitative PCR method to detect white spot syndrome virus in shrimp, but did not prepare a standard curve, so the real quantitative detection has not yet been achieved.

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