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Duck hepatitis bivalent live vaccines and preparation method thereof

A live vaccine, duck hepatitis technology, applied in the field of duck hepatitis bivalent live vaccine and its preparation

Active Publication Date: 2016-07-06
QILU ANIMAL HEALTH PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the only live vaccine used to prevent duck hepatitis in the domestic market is duck hepatitis virus serotype 1 monovalent vaccine

Method used

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  • Duck hepatitis bivalent live vaccines and preparation method thereof
  • Duck hepatitis bivalent live vaccines and preparation method thereof
  • Duck hepatitis bivalent live vaccines and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] - Inspection of poisonous species

[0080] 1 Sterility test and mycoplasma test

[0081]DHV-1QL1 strain virus seeds were tested for sterility and mycoplasma according to the appendix of the current "Chinese Veterinary Pharmacopoeia", and there was no contamination by bacteria, mold and mycoplasma.

[0082] DHV-3QL3 strains were tested for sterility and mycoplasma according to the appendix of the current "Chinese Veterinary Pharmacopoeia", and there was no contamination by bacteria, mold and mycoplasma.

[0083] 2 Exogenous virus inspection

[0084] DHV-1QL1 strain virus seeds are tested for exogenous virus by chicken embryo culture, cell culture and ELISA test (avian leukemia virus detection) according to the appendix of the current "Chinese Veterinary Pharmacopoeia", and there is no exogenous virus contamination.

[0085] The DHV-3QL3 strain virus species was tested for exogenous virus by chicken embryo culture, cell culture and ELISA test (detection of avian leukosi...

Embodiment 2

[0102] ——The virulence recovery test of the poisonous species

[0103] 1 material

[0104] 1.1 Strain type 1 duck hepatitis virus QL1 strain virus seed, virus content is 10 8.38 ELD 50 / 0.1ml; type 3 duck hepatitis virus QL3 strain virus seed, virus content is 10 7.68 ELD 50 / 0.1ml.

[0105] 1. 28-10-day-old SPF chicken embryos were purchased from Jinan Spafas Poultry Co., Ltd.

[0106] 1. The 31-day-old healthy susceptible ducks came from healthy breeding ducks with no history of duck hepatitis and negative DHAV antibodies (neutralizing antibody titer <1:4).

[0107] 2 methods

[0108] 2.1 Virulence recovery test of DHV-1QL1 strain

[0109] 2.1.1 Inoculation of 1-day-old healthy susceptible ducks Inoculate 5 1-day-old healthy susceptible ducks with type 1 duck hepatitis virus QL1 strain virus seed by intramuscular injection, 0.5ml / piece. Observe and record the clinical manifestations of ducklings such as spirit, diet, and feces every day after inoculation, and dissect...

Embodiment 3

[0134] - vaccine preparation

[0135] 1 poison seed breeding

[0136] Take DHV-1QL1 strain virus seeds, dilute 100 times with sterilized normal saline, inoculate 9-10 day-old SPF chicken embryos through the allantoic cavity route, 0.2ml per embryo, incubate at 37°C, discard dead embryos within 24 hours, Aseptically collect the allantoic fluid of chicken embryos that died within 36 to 96 hours, and store it at -20°C. At the same time, do a sterility test and determine the virus content. The measured virus content should be ≥ 10 9.0 ELD 50 / ml. The virus allantoic fluid that has passed the sterility test and the virus content determination is used as the virus species for production.

[0137] Take DHV-3QL3 strain virus seeds, dilute 100 times with sterilized normal saline, inoculate 8-10 day-old SPF chicken embryos through the chorioallantoic membrane route or inoculate 6-7 day-old SPF chicken embryos through the yolk sac route, 0.2ml per embryo , incubated at 37°C, discarde...

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Abstract

The invention relates to duck hepatitis bivalent live vaccines and a preparation method thereof. According to the invention, 1) SPF chicken embryos are inoculated with vaccine strains of the duck hepatitis bivalent live vaccines, namely DHV-1 QL1 strains, through allantoic cavities for culture, and are inoculated with DHV-3 QL3 strains through chorioallantoic membranes or yolk sacs for culture, the antigen contents of the prepared vaccines are high, and all not less than 10<8.5>ELD50 / ml, so that the duck hepatitis bivalent live vaccines have the characteristics of good safety and excellent immunogenicity, and break through limitations of DHV-3 type virus strains which only can be proliferated in duck embryos; 2) virus solutions cultured by the SPF chicken embryos lower risks of exogenous pathogenic microbial contamination, improve labor efficiency and reduce production costs, thereby being suitable for mass production; 3) the bivalent live vaccines disclosed by the invention can prevent DHV-1 type and DHV-3 type virulent viruses attacking on ducklings; 4) the duck hepatitis bivalent live vaccines are simple in production process, easy in quality control and small in batch difference, thereby providing a guarantee for producing safe and qualified vaccines.

Description

technical field [0001] The invention relates to a bivalent live vaccine for duck hepatitis and a preparation method thereof. It belongs to the field of veterinary biological products. Background technique [0002] Duck hepatitis (Duck Hepatitis, DH) is caused by duck hepatitis virus (Duck Hepatitis virus, DHV), a kind of hemorrhagic inflammation of the liver, which is an acute, highly contagious and fatal infectious disease of ducklings. The disease can occur throughout the year, and only occurs in ducklings during natural outbreaks, causing huge economic losses to the breeding industry. Traditional duck hepatitis virus has three serotypes, which cause type 1, type 2 and type 3 duck hepatitis respectively (Lu Chengping. Veterinary Microbiology (Third Edition) [M]. Beijing: China Agricultural Press, 2001, 564). In recent years, due to immune pressure and other factors, serotype Ⅰ duck hepatitis virus presents a new epidemic situation. Scholars in Taiwan, China and South Ko...

Claims

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Application Information

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IPC IPC(8): A61K39/125A61P31/14C12N7/00C12R1/93
CPCA61K39/12A61K2039/5254A61K2039/70C12N7/00C12N2770/32334C12N2770/32351
Inventor 王蕾张青婵刘旭光吴昊张中波房宝英徐龙涛
Owner QILU ANIMAL HEALTH PROD
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