A kind of attenuated strain of carp herpesvirus type II and its application

A carp herpes virus and attenuated strain technology, applied in the direction of viruses, antiviral agents, viruses/bacteriophages, etc., can solve the problems of high lethality, strong infectivity, and economic losses in crucian carp breeding, so as to improve the survival rate and reduce the incidence of disease rate effect

Active Publication Date: 2021-03-23
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Crucian carp hematopoietic organ necrosis caused by cyprinid herpesvirus 2 (CyHV-2) infection in cultured crucian carp is a newly discovered viral disease infecting cultured crucian carp in my country. Crucian carp farming causes huge economic losses

Method used

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  • A kind of attenuated strain of carp herpesvirus type II and its application
  • A kind of attenuated strain of carp herpesvirus type II and its application
  • A kind of attenuated strain of carp herpesvirus type II and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Isolation and identification of carp herpesvirus type II CyHV-2DX2019

[0020] (1) Detection of carp herpesvirus type II

[0021] The carp herpes virus type II CyHV-2DX2019 attenuated strain of the present invention is taken from silver carp in a crucian carp-silver carp polyculture pond in Jingzhou, Hubei. The silver carp kidney tissue is placed in a sterile culture dish and cut with sterile ophthalmic scissors Add 10 times the volume (V / W) of phosphate buffered solution (PBS), transfer to a glass homogenizer, and grind into a tissue homogenate under ice-bath conditions. The DNA of tissue homogenate was extracted and amplified with primers CyHV2Hel-F: 5’-GGACTTGCGAAGAGTTTGATTTCTTAC-3’ and CyHV2Hel-R: 5’-CCATAGTCACCATCGTCTCATC-3’. The gel running results of the amplified product showed that the test sample had a specific band at 360 bp, which was consistent with the band size of the positive result of carp herpesvirus type II. After the amplified product was sequenced...

Embodiment 2

[0030] Pathogenicity experiment of CyHV-2DX2019 strain of carp herpesvirus type II to crucian carp

[0031] Take 180 crucian carp, with an average body weight of (150.0±4.0) g and an average body length of (25.0±2.5) cm. Before the experiment, crucian carp needs to be tested for viral nucleic acid to confirm that the carp herpesvirus type II is negative before it can be used in the experiment. Crucian carp is randomly divided into 6 groups according to the test strain, with 30 tails in each group, and the fresh culture of the 6th-10th generation test strain (CyHV-2DX2019 strain) is inoculated respectively by intraperitoneal injection method, 0.2ml / tail, the virus concentration of the virus liquid ≥5×10 5 copies / ml, the control group was inoculated with PBS in the same way, and observed continuously for 14 days.

[0032]The results showed that the behavior, appetite and other physiological indicators of the crucian carp inoculated with different generations of viruses were no...

Embodiment 3

[0034] Preparation of live vaccine against hematopoietic organ necrosis of crucian carp

[0035] In Example 1, the 10th and subsequent generations of inoculated cells blindly passed were observed continuously for 14 days, and then the cell cultures were harvested. After being frozen at -80°C, it was thawed at room temperature, repeated freezing and thawing three times, and then centrifuged at 4000r / min for 30min at 4°C. The supernatant was taken to extract virus DNA, and the virus content was detected by digital PCR. The virus concentration ≥ 5 × 10 5 The supernatant of copies / ml was diluted 100 times with PBS to prepare a live vaccine for crucian carp hematopoietic organ necrosis. 3 copies / ml.

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Abstract

The invention belongs to the technical field of vaccine preparation, and particularly provides a cyprinid herpesvirus 2 attenuated strain and an application thereof. The cyprinid herpesvirus 2 provided by the invention is a cyprinid herpesvirus 2 DX2019 strain, and the preservation number is CCTCC NO:V201980. A vaccine prepared by using the strain can prevent diseases caused by the cyprinid herpesvirus 2, has an immune protection effect of 92 percent or above, does not generate a stress reaction to organisms, and can be applied to prevention of haematopoietic organ necrosis of crucian carps, so that the incidence rate of haematopoietic organ necrosis of the crucian carps in a crucian carp culture pond is reduced, and the survival rate of the crucian carps is improved.

Description

technical field [0001] The invention belongs to the technical field of vaccine preparation, in particular to a type II attenuated carp herpes virus strain and application thereof. Background technique [0002] Crucian carp is an important freshwater aquaculture species in my country. According to statistics, the total output of crucian carp in China in 2018 was 2.77 million tons. However, with the gradual expansion of the scale of crucian carp farming, the continuous improvement of the degree of intensification and the continuous deterioration of the aquaculture water environment, the problem of crucian carp disease has become increasingly prominent, causing major economic losses to the aquaculture industry. At present, viral diseases have become one of the main factors threatening the healthy development of crucian carp aquaculture in my country. [0003] Crucian carp hematopoietic organ necrosis caused by cyprinid herpesvirus 2 (CyHV-2) infection in cultured crucian carp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/245A61P31/22C12R1/93
CPCA61K39/12A61K2039/5254A61K2039/545A61K2039/552A61P31/22C12N7/00C12N2710/16021C12N2710/16034
Inventor 周勇薛明洋范玉顶刘文枝孟彦江南李逸群曾令兵
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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