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.
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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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Abstract
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