DHAV (Duck Hepatitis A Virus) III type complex live vaccine and preparation method thereof

A duck hepatitis A virus and complex technology, applied in biochemical equipment and methods, vaccines, viruses, etc., can solve the problems of time-consuming, labor-intensive, expensive vaccines, and high production costs, and achieve a wide safety range, good safety, and good protection. effect of action

Active Publication Date: 2017-04-19
哈药集团生物疫苗有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The production method of the traditional antigen-antibody complex vaccine is to combine the hyperimmune serum with the virus. T

Method used

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  • DHAV (Duck Hepatitis A Virus) III type complex live vaccine and preparation method thereof
  • DHAV (Duck Hepatitis A Virus) III type complex live vaccine and preparation method thereof
  • DHAV (Duck Hepatitis A Virus) III type complex live vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Virus and egg yolk antibody ratio screening and detection test

[0028] 1. Preparation of virus stock solution

[0029] 1.1 Inoculation

[0030] Duck hepatitis A virus type III JS57 strain (microorganism preservation number is: CGMCC No.9708) is made 1000-fold dilution with sterilized normal saline, inoculates 0.2ml in every embryo allantoic cavity of 9-day-old SPF chicken embryo, seals pinhole after inoculation, Continue incubation at 37°C without turning the eggs.

[0031] 1.2 Incubation and observation

[0032] After 24 hours of inoculation, the eggs were illuminated every 4 hours until 96 hours. Take chicken embryos that died within 24 to 96 hours and cool them at 2 to 8°C.

[0033] 1.3 harvest

[0034] Chick embryos that have been cooled for 4 to 24 hours are collected by aseptic surgery, including chorioallantoic membrane, fetus, embryo fluid and amniotic fluid, and each of them is mixed into a group and placed in a sterilized bottle. While harvest...

Embodiment 2

[0072] Example 2 Preparation of duck hepatitis A virus type III complex live vaccine

[0073] 1. Preparation of virus stock solution

[0074] 1.1. Preparation of poisonous seeds for production

[0075] Take 9-day-old SPF chicken embryos, inoculate the basic virus species (that is, duck hepatitis A virus type Ⅲ chicken embryo attenuated strain JS57 strain (microorganism preservation number: CGMCC No.9708)), incubate at 37°C, and irradiate the embryos 2 to 4 times a day , Observed for 4 days, discarded dead chicken embryos within 24 hours, and collected allantoic fluid and embryo bodies from chicken embryos that died within 96 hours for sterility testing, and observed fetal lesions. Freezing and thawing 3 times, centrifuging at 4,000rpm for 30min, absorbing the supernatant, quantitatively dispensing into glass bottles, storing in a -80°C refrigerator, and inspecting according to the inspection standards specified by the production seed batch, to obtain the production poisonous ...

Embodiment 3

[0122] Freeze and vacuum dry immediately after subpackaging, according to page 437 of the appendix of the 2000 edition of the "Procedures". Example 3 Duck Hepatitis A Virus Type III Complex Live Vaccine Finished Product Inspection

[0123] The finished product of duck hepatitis A virus type III complex live vaccine prepared in Example 2 of the present invention was tested.

[0124] 1. Physical properties

[0125] It is a light red spongy loose mass, easy to separate from the bottle wall, and dissolves quickly after adding diluent.

[0126] 2. Sterility test

[0127] Tested according to the appendix of "Chinese Veterinary Pharmacopoeia", it should grow aseptically.

[0128] 3. Mycoplasma test

[0129] Tested according to the appendix of "Chinese Veterinary Pharmacopoeia", there should be no mycoplasma growth.

[0130] 4. Exogenous virus inspection

[0131] Test according to the appendix of "Chinese Veterinary Pharmacopoeia", there should be no exogenous virus contaminatio...

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Abstract

The invention discloses a DHAV (Duck Hepatitis A Virus) III type complex live vaccine and a preparation method thereof. The method firstly discloses the DHAV III type complex live vaccine, which comprises antigen-antibody complex and pharmaceutically acceptable freeze-drying protective agents, wherein the antigen-antibody complex is prepared from DHAV III type chick embryo weakening strain JS57 virus liquid and duck anti-DHAV III type egg yolk antibodies. The invention further discloses a preparation method of the DHAV III type complex live vaccine. The preparation method comprises the following steps of mixing the freeze-drying protective agents with the antigen-antibody complex prepared from the DHAV III type chick embryo weakening strain JS57 virus liquid and the duck anti-DHAV III type egg yolk antibodies; performing freeze vacuum drying; and obtaining the DHAV III type complex live vaccine. The preparation method provided by the invention has the advantages that the antibody obtaining path is simple; the production cost is low; the prepared DHAV III type complex live vaccine can break through the maternal antibody interference to generate high neutralizing antibody valence; and the production rate on ducklings is improved.

Description

technical field [0001] The invention relates to a duck hepatitis A virus type III complex live vaccine, and also relates to a preparation method of duck hepatitis A virus type III complex live vaccine, which belongs to the field of preparation of duck hepatitis A virus type III antigen-antibody complex vaccine. Background technique [0002] Duck hepatitis A virus (DHAV) belongs to the Picornaviridae family and the genus Avian Hepadnavirus. At present, DHAV is divided into three serotypes, namely: serotype I, serotype Ⅱ and serotype Ⅲ. Among them, serotype I is the traditional duck hepatitis virus, and serotypes Ⅱ and Ⅲ are new virus strains discovered in recent years, corresponding to the new strains from Taiwan and South Korea respectively, and there is no cross-reaction between the 3 serotypes. [0003] Duck viral hepatitis is an acute contact infectious disease of ducklings caused by duck hepatitis A virus, which mainly harms ducklings under 3 weeks old, with a mortality...

Claims

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Application Information

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IPC IPC(8): A61K39/42A61K39/29A61P1/16A61P31/14
CPCA61K39/12A61K39/42A61K2039/545A61K2039/552A61K2039/57C12N2770/32334A61K2300/00
Inventor 藏玉婷高艳春丁国杰陈欣王彬李立莉郑铁鑫尹尧秦立鑫时鹏飞张凤强
Owner 哈药集团生物疫苗有限公司
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