Method for establishing prawn virus isolation, identification and infection model

A technology for virus isolation and model establishment, applied in the field of anti-virus research, it can solve problems that have not yet been reported, and achieve the effect of clear judgment criteria, obvious infection effect, and good reproducibility.

Inactive Publication Date: 2013-03-20
GUANGXI INST OF FISHERIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, only the public literature on the establishment of the shrimp white spot syndrome virus infection model has been found, but there has been no report on the establishment of the shrimp taura virus (TSV) infection model

Method used

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  • Method for establishing prawn virus isolation, identification and infection model
  • Method for establishing prawn virus isolation, identification and infection model
  • Method for establishing prawn virus isolation, identification and infection model

Examples

Experimental program
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Effect test

Embodiment 1

[0020] A method for isolation, identification and infection model establishment of shrimp virus, comprising the following steps:

[0021] (1) Obtainment of TSV virus tissue: The preserved TSV purified virus particles were diluted at a mass-to-volume ratio of 1:1000, and injected into the muscle of the third abdominal ganglion of Litopenaeus vannamei with an insulin syringe. ~12g), inject 10~12μL per tail (inject the virus at a dose of 1μL / g to each prawn), control the shrimp culture water temperature at 28-30℃, salinity 28‰ after injection, continuously aerate, observe the status of the prawns every day, Collect the dying prawns in time and place them in liquid nitrogen;

[0022] (2) TSV virus detection: PCR was used to detect TSV virus infection of shrimp, and RNA from hepatopancreatic tissue of dying shrimp was extracted, reverse transcribed into cDNA, and amplified using this cDNA as a template to amplify the upstream and downstream primer sequences of TSV virus. Upstream ...

Embodiment 2

[0028] A method for isolation, identification and infection model establishment of shrimp virus, comprising the following steps:

[0029] (1) Obtainment of TSV virus tissue: The preserved TSV purified virus particles were diluted at a mass-to-volume ratio of 1:1000, and injected into the muscle of the third abdominal ganglion of Litopenaeus vannamei with an insulin syringe. ~12g), inject 10~12μL per tail (inject the virus at a dose of 1μL / g to each prawn), control the shrimp culture water temperature at 28-30℃, salinity 28‰ after injection, continuously aerate, observe the status of the prawns every day, Collect the dying prawns in time and place them in liquid nitrogen;

[0030](2) TSV virus detection: PCR was used to detect TSV virus infection of shrimp, and RNA from hepatopancreatic tissue of dying shrimp was extracted, reverse transcribed into cDNA, and amplified using this cDNA as a template to amplify the upstream and downstream primer sequences of TSV virus. Upstream p...

Embodiment 3

[0036] A method for isolation, identification and infection model establishment of shrimp virus, comprising the following steps:

[0037] (1) Obtainment of TSV virus tissue: The preserved TSV purified virus particles were diluted at a mass-to-volume ratio of 1:1000, and injected into the muscle of the third abdominal ganglion of Litopenaeus vannamei with an insulin syringe. ~12g), inject 10~12μL per tail (inject the virus at a dose of 1μL / g to each prawn), control the shrimp culture water temperature at 28-30℃, salinity 28‰ after injection, continuously aerate, observe the status of the prawns every day, Collect the dying prawns in time and place them in liquid nitrogen;

[0038] (2) TSV virus detection: PCR was used to detect TSV virus infection of shrimp, and RNA from hepatopancreatic tissue of dying shrimp was extracted, reverse transcribed into cDNA, and amplified using this cDNA as a template to amplify the upstream and downstream primer sequences of TSV virus. Upstream ...

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PUM

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Abstract

The invention discloses a method for establishing a prawn virus isolation, identification and infection model, which is characterized by comprising the following steps: (1) obtaining TSV (Taura Syndrome Virus) tissues; (2) detecting TSV viruses; (3) preparing TSV fluid; and (4) building a TSV infection model. A prawn TSV virus infection model established by using the method disclosed by the invention is good in reproducibility, the virulence of RNA (Ribonucleic Acid) viruses is relatively strong, and the infection effect is obvious; and an identification method is simple, and clear in judgment standards, not only applicable to the research on the antiviral breeding of RNA viruses, but also capable of being applied to the research on the TSV pathogenesis.

Description

technical field [0001] The invention belongs to the field of anti-virus research, in particular to a method for isolating, identifying and establishing infection models of prawn viruses. Background technique [0002] Litopenaeus vannamei has been introduced into my country for large-scale farming because of its fast growth, strong disease resistance, wide salt tolerance, and delicious meat taste. At present, 80% of the shrimp cultured in my country are Litopenaeus vannamei. The breeding process is also accompanied by the massive reproduction and spread of viral diseases. The outbreak of viral diseases is still the biggest obstacle to the healthy development of the shrimp farming industry. Among them, Taura Syndrome Virus (TSV) is the One of the most serious pathogens in aquaculture. TSV virus is a single-stranded RNA virus. The main target organs of infection are crustacean epithelium (appendix, gills, stomach, esophagus, hindgut), connective tissue, etc., causing shrimp dise...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12Q1/70C12Q1/68A01K67/033C12R1/93
Inventor 陈秀荔赵永贞陈晓汉彭敏李咏梅杨春玲何苹萍
Owner GUANGXI INST OF FISHERIES
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