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Inactivated vaccine for duck viral hepatitis as well as preparation method and application thereof

A technology for duck viral hepatitis and duck hepatitis A, which is applied in the field of inactivated duck viral hepatitis vaccine and its preparation, can solve the problems of susceptibility to virulence, long survival time, immunization failure, etc., and achieves the duration of immunization. Long, stable immune effect, easy to use effect

Active Publication Date: 2015-04-22
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The attenuated live vaccine has the following problems: (1) storage and transportation require low temperature conditions of -20°C, which is inconvenient; (2) after immunizing ducks, the vaccine virus survives in the body for a long time and is easy to detoxify, and the virus has strong resistance to the environment and is easy to detoxify. (3) The immune effect of the vaccine is unstable and easily interfered by maternal antibodies, resulting in immune failure

Method used

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  • Inactivated vaccine for duck viral hepatitis as well as preparation method and application thereof
  • Inactivated vaccine for duck viral hepatitis as well as preparation method and application thereof
  • Inactivated vaccine for duck viral hepatitis as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Embodiment 1, the acquisition of DHAV-SH strain

[0058] 1. Discovery of DHAV-SH strain

[0059] In a duck farm in Shanghai in December 2013, Liu Wenjun and Li Jing isolated a strain of duck viral hepatitis type Ⅰ virus from the diseased duck, and named it DHAV-SH strain.

[0060] 2. Identification of DHAV-SH strain

[0061] 1. Virus form

[0062] Virus particles of the DHAV-SH strain are icosahedral and about 40 nm in diameter.

[0063] 2. Molecular biological identification

[0064] Extract the RNA of the DHAV-SH strain, obtain the DNA after reverse transcription, design a pair of specific primers (P1 and P2) according to the DHAV complete gene sequence published by GENBANK, and use P1 and P2 to perform RT-PCR amplification on the obtained DNA , to obtain the amplified product. Sequencing of the amplified product showed that the target fragment with a size of 229 bp was amplified, and its sequence is shown in sequence 1 of the sequence list, which has a high homo...

Embodiment 2

[0082] Embodiment 2, the preparation of duck viral hepatitis bivalent inactivated vaccine

[0083] 1. Antigen preparation

[0084] 1. Preparation of virus liquid

[0085] The DHAV-SH strain was inoculated into 11-day-old susceptible duck embryos through the allantoic cavity, 0.1 mL per embryo, incubated at 37°C for 120 hours, then aseptically collected embryo liquid and embryo body, homogenized at high speed (14000r / min), and then used Filter through 8 layers of sterile gauze, collect the filtrate, and freeze and thaw once at -20°C to obtain the DHAV-SH strain virus liquid. Virus content=10 of the DHAV-SH strain virus liquid of every 0.1mL 6.25 ELD 50 (Duck embryo half lethal dose). In actual application, the virus content of every 0.2mL of DHAV-SH strain virus liquid is ≥10 6.25 ELD 50 can be.

[0086] The DHAV-FS strain was inoculated into 11-day-old susceptible duck embryos through the allantoic cavity, 0.1 mL per embryo, and incubated at 37°C for 120 hours, then ase...

Embodiment 3

[0097] Embodiment 3, the stability of duck viral hepatitis bivalent inactivated vaccine

[0098] Take 3 batches of duck viral hepatitis bivalent inactivated vaccine and store them at 4°C. After 3, 6, 9, 12, and 15 months of storage, take samples for physical property testing, and compare the test results with those after production .

[0099] The results showed that after 3, 6, 9, 12, and 15 months of storage at 2-8°C, the physical properties of the three batches of laboratory products remained stable, and the appearance, dosage form, stability, and viscosity all conformed to the "Trial Regulations" (draft) ) in the quality standards. See Table 1 for detailed results.

[0100] Table 1. Test results of physical properties of 3 batches of laboratory products at different storage times

[0101]

[0102]

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Abstract

The invention discloses an inactivated vaccine for duck viral hepatitis as well as a preparation method and an application thereof. The bivalent inactivated vaccine for duck viral hepatitis comprises an antigen and an oil adjuvant, wherein the antigen is prepared by mixing a duck hepatitis viral composition and Tween-80; the duck hepatitis viral composition comprises a duck hepatitis A virus (DHAV) type 1 and a duck hepatitis A virus (DHAV) type 3; the duck hepatitis A virus type 1 is an inactivated duck hepatitis virus type 1 DHAV-SH strain virus liquid; the duck hepatitis A virus type 3 is an inactivated duck hepatitis virus type 3 DHAV-FS strain virus liquid; and the oil adjuvant comprises Montanide ISA206, aluminum stearate and span-80. The bivalent inactivated vaccine for duck viral hepatitis, which is disclosed by the invention, has the following advantages that (1) the inactivated vaccine simultaneously has preventive effects on the duck hepatitis A virus type 1 and the duck hepatitis A virus type 3; (2) the inactivated vaccine is prepared from inactivated virus and has good safety; (3) the inactivated vaccine is convenient to store, transport and use; and (4) the stable immune effect is achieved.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to an inactivated vaccine of duck viral hepatitis, a preparation method and application thereof. Background technique [0002] Duck viral hepatitis, referred to as duck hepatitis for short, is an acute severe infectious disease characterized by hemorrhagic inflammation in the liver caused by duck viral hepatitis virus. Sick ducks and infected ducks are the main sources of infection, mainly through the digestive tract and respiratory tract. Poor feeding management, lack of vitamins and minerals, damp and crowded duck houses can all promote the occurrence of duck viral hepatitis. Once duck viral hepatitis occurs, it spreads quickly among ducks, and the incidence rate can reach 100%. Duck viral hepatitis mainly infects ducklings within 20 days of age, with an incubation period of 1-4 days, sudden onset and short course of disease. At the beginning ...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/29A61K39/39A61P31/14C12R1/93
Inventor 刘文军李晶张爽张万林范国兵
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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