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A kind of largemouth bass iridovirus inactivated vaccine for injection and preparation method thereof

A virus inactivation and inactivation vaccine technology, applied in veterinary vaccines, biochemical equipment and methods, viruses, etc., can solve the problems of outbreaks of diseases, economic losses, etc., achieve good immunogenicity, prevent the occurrence and course of infection The effect of the progress

Active Publication Date: 2022-07-29
NANYANG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

From 2008 to the beginning of 2010, largemouth bass cultured fishponds in Shunde area suffered from large-scale disease outbreaks, causing heavy economic losses
However, at present, there is no specific drug for the prevention and treatment of iridescent virus, and there is no commercial iridescent virus vaccine in my country. Therefore, isolating suitable vaccine strains and carrying out related research to promote the industrial production of vaccines is very important for the breeding industry of largemouth bass. It has positive significance and is also an urgent industrial problem to be solved

Method used

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  • A kind of largemouth bass iridovirus inactivated vaccine for injection and preparation method thereof
  • A kind of largemouth bass iridovirus inactivated vaccine for injection and preparation method thereof
  • A kind of largemouth bass iridovirus inactivated vaccine for injection and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. Isolation and identification of largemouth bass iridovirus

[0035] 1. Collection of largemouth bass iridovirus-infected samples

[0036] From June to July 2019, a large-scale disease of largemouth bass broke out in a farm in Nanyang City, Henan Province. The main symptoms were black body color, lack of vitality, mild abdominal swelling, mortality rate as high as 70%, and dying fish. After necropsy, symptoms of acute peritonitis, necrosis and inflammatory lesions appeared in the surface part of liver, stomach, intestine and spleen, but the deep part appeared normal, there was fibrin exudate on the contact surface of swim bladder and abdominal cavity, mucosa of gastrointestinal tract There were localized necrotic lesions in the epithelium, which were in line with the symptoms associated with largemouth bass iridovirus infection. Twenty largemouth bass with ulcer symptoms were collected from the farm. The collected samples were stored separately in sealed bags...

Embodiment 2

[0055] Example 2. The preparation method of largemouth bass iris virus N-LBIV-201907 inactivated vaccine

[0056] Step 1, culture of EPC cells: subculture EPC cells in M199 medium containing 10% (V / V) fetal bovine serum at 25°C, 5% CO 2 Incubate at constant temperature until the cells are confluent, grow into a monolayer, and the number of cells is > 10 6 / mL

[0057] Step 2, Proliferation of Largemouth Bass Iridescent Virus N-LBIV-201907: Aspirate the culture medium in the EPC cells, and then add the virus suspension of N-LBIV-201907 (10 5 TCID 50 / mL), after the virus was adsorbed to the cells for 1 h, M199 (pH 7.2-7.5) maintenance solution containing 2% (V / V) calf serum was added for virus proliferation.

[0058] Step 3: Virus harvesting: The virus is continuously cultured for 4 to 7 days, observed under a microscope, and when 80% of the EPC cells show typical cytopathic effects, they are placed in a -80°C refrigerator for freezing, and after freezing, they are taken out...

Embodiment 3

[0065] Example 3. Safety inspection and immune efficacy evaluation of largemouth iridovirus inactivated vaccine

[0066] 1. Safety inspection of inactivated vaccines

[0067] 1.1 Inactivation effect test

[0068] Take the prepared vaccine and inoculate EPC cells according to the above method of virus proliferation, and set a negative control (freezing and thawing supernatant of EPC cells without virus) at the same time, observe for 7 days, if cytopathic effect occurs, it means that the virus inactivation is incomplete; If no cytopathic effect is seen, blind pass 3 more times. If cytopathic occurs in blind pass, it means that virus inactivation is still incomplete.

[0069] Sterility testing

[0070] Take the vaccine prepared above, inoculate the brain infusion bacterial culture medium (BHI) plate, spread the plate with streak method and culture at 37℃ for 15 days. Use after membrane filtration sterilization; if no colonies grow, the vaccine is sterile.

[0071] fish safety...

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Abstract

The present invention aims to provide an inactivated largemouth bass iridovirus vaccine for injection and a preparation method thereof. Based on a newly isolated largemouth bass iridovirus, after inactivation, it is mixed with an adjuvant as an inactivated vaccine. It shows that the prepared inactivated vaccine has good immune effect, and the relative immune protection rate can reach up to 100%, which can be widely used in the prevention and control of iridovirus infection in largemouth bass.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a largemouth bass iridovirus inactivated vaccine for injection and a preparation method thereof. Background technique: [0002] largemouth bass ( Micropterus salmoides ), commonly known as California perch and black perch, native to the Mississippi River Basin in North America, is a wide-temperature fish, with many advantages such as fast growth, less disease, delicious meat, high price and easy fishing, and is deeply loved by fishermen. In the 1970s, largemouth bass was first introduced to Taiwan, China. After more than ten generations of breeding, it was introduced to Guangdong Province in 1983 and artificially cultured in Shenzhen, Foshan and Huizhou. The Shunde area of ​​Foshan is the main breeding area for largemouth bass. Because the breeding of largemouth bass has good economic benefits in China, it has rapidly developed into the main economic fish species in my ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/12A61P31/20C12N7/00C12N7/06
CPCA61K39/12A61P31/20C12N7/00A61K2039/54A61K2039/5252A61K2039/552C12N2710/00034C12N2710/00052C12N2710/00063
Inventor 姚伦广孟晓红邱礽秦英惠管翠翠刘阳坤
Owner NANYANG NORMAL UNIV
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