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Inactivated vaccine for type 2 rabbit hemorrhagic disease virus and preparation method thereof

A technology of rabbit hemorrhagic virus and inactivated vaccine, which is applied in the field of type 2 rabbit hemorrhagic virus inactivated vaccine and its preparation, can solve the problems that RHDV2 infection cannot produce effective protection, genetic characteristics and antigenic differences, etc., and achieves safety High performance, good immune effect and simple process

Active Publication Date: 2020-09-22
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

RHDV2 and classic RHDV strains infect dead rabbits with similar symptoms and pathological changes, but there are great differences in genetic characteristics and antigenicity, and the vaccine prepared by classic strains cannot produce effective protection against RHDV2 infection

Method used

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  • Inactivated vaccine for type 2 rabbit hemorrhagic disease virus and preparation method thereof
  • Inactivated vaccine for type 2 rabbit hemorrhagic disease virus and preparation method thereof
  • Inactivated vaccine for type 2 rabbit hemorrhagic disease virus and preparation method thereof

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Experimental program
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Effect test

preparation example Construction

[0022] The invention provides a preparation method of an inactivated vaccine of type 2 rabbit hemorrhagic virus, comprising the following steps: 1) inoculating susceptible rabbits with type 2 rabbit hemorrhagic virus strain SC2020; 2) collecting within 24-96 hours after the inoculation Organ tissue of a dead rabbit with typical histopathological changes; 3) Mix the organ tissue with normal saline at a ratio of 1 g:(4-6) mL, homogenize and filter to obtain tissue homogenate; 4) The tissue homogenate is mixed with physiological saline until the ratio of organ tissue to physiological saline is 1 g: (18-20) mL to obtain vaccine antigens, and the vaccine antigens are inactivated to obtain type 2 rabbit hemorrhagic virus inactivated vaccine .

[0023] In the present invention, susceptible rabbits were inoculated with type 2 rabbit hemorrhagic virus strain SC2020. In the present invention, the type 2 rabbit hemorrhagic virus strain SC2020 is isolated from Sichuan, China, and the Gen...

Embodiment 1

[0031] strain screening

[0032] The liver samples of commercial rabbits and young rabbits from 2 different regions that have been diagnosed were removed, the connective tissue on them was removed, and the normal saline was added to smash and filter to prepare a tissue homogenate. The homogenate contained the mashed liver weight (g): The volume (ml) of normal saline was 1:9, and the freeze-thaw was repeated 3 times. Centrifuge at 8000rpm for 30min, take the supernatant and filter it with a 0.22μm filter, respectively numbered JT1, JT2, ZY1, ZY2, and inoculated subcutaneously into healthy susceptible rabbits aged 2 to 4 months, 3 rabbits / group, 1 mL / Only. The dead rabbits were necropsied after inoculation, and the liver tissue was aseptically collected and placed in a sterile container. The homogenate was diluted 1:9 with physiological saline, and the hemagglutination titer was determined by repeated freezing and thawing 3 times. The results are shown in Table 1 below.

[00...

Embodiment 2

[0038] Subculture of strains

[0039] The SC2020 / 04-F1 was centrifuged at 8000 rpm for 30 min, and the supernatant was taken and sterilized with a 0.22 μm filter, labeled as R2-F1. R2-F1 was subcutaneously inoculated into 3 healthy susceptible rabbits aged 2-4 months, 1 mL / rabbit. Necropsy of dead rabbits after inoculation, aseptically collected rabbit livers that died within 24-96 hours after inoculation, and the death time did not exceed 1 hour with histopathological changes as virus seeds, put them in sterile containers, and removed the connective tissue on them. The homogenate was diluted 1:9 with normal saline, freeze-thawed 3 times, labeled as SC2020 / 04-F2, and cryopreserved. According to the above method, the virus strain was inoculated with healthy susceptible rabbits for continuous passage to the 8th generation, and the hemagglutination titer was measured, and then cryopreserved respectively. The results are shown in Table 2 below.

[0040]Table 2 Results of hemaggl...

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Abstract

The invention provides an inactivated vaccine for type 2 rabbit hemorrhagic disease virus and a preparation method thereof, and belongs to the technical field of vaccine preparation. The preparation method comprises the following steps: 1) inoculating a susceptible rabbit with a type 2 rabbit hemorrhagic disease virus strain SC2020; 2) collecting organ tissues of rabbits which die within 24-96 hours after inoculation and have typical tissue pathological changes; 3) mixing the organ tissues with normal saline according to a ratio of 1g:(4-6)mL, and performing homogenizing and filtering to obtain tissue homogenate; and 4) mixing the tissue homogenate with normal saline until the ratio of the organ tissues to the normal saline is 1g:(18-20)mL to obtain a vaccine antigen, and inactivating thevaccine antigen to obtain the inactivated vaccine for the type 2 rabbit hemorrhagic disease virus. The inactivated vaccine for the type 2 rabbit hemorrhagic disease virus is high in safety, good in immune effect and simple and convenient in process.

Description

technical field [0001] The invention belongs to the technical field of vaccine preparation, and in particular relates to a type 2 rabbit hemorrhagic virus inactivated vaccine and a preparation method thereof. Background technique [0002] Rabbithemorrhagic disease (RHD) is an acute, highly contagious and highly lethal disease caused by rabbit hemorrhagic disease virus (RHDV). Classic RHDV only infects rabbits, and rabbits over 2 months old are susceptible, and the mortality rate is as high as 90%, often 48 to 72 hours after infection. The dead rabbits are caused by blood stasis in the respiratory system and parenchymal organs such as liver, spleen, kidney, and heart. , swelling and hemorrhage are characterized, and young rabbits are not susceptible. In 2010, a new strain of RHDV, named RHDV2, was first discovered in France. Rabbits of different ages are susceptible to RHDV2, and the mortality rate of infected rabbits can reach 90%. The symptoms and pathological changes of...

Claims

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Application Information

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IPC IPC(8): A61K39/12A61P31/14
CPCA61K39/12A61P31/14A61K2039/552A61K2039/5252C12N2770/16034Y02A50/30
Inventor 范志宇王芳胡波魏后军陈萌萌仇汝龙宋艳华朱伟峰薛家宾
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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