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DHAV (duck hepatitis A virus) JS strain and application of DHAV JS strain in duck virus hepatitis prevention and cure

A technology of duck hepatitis A virus and microbial strains, which is applied in the direction of antiviral agents, viruses/phages, and medical preparations containing active ingredients, to achieve the effects of good safety, improved production performance, and reduced injection doses

Active Publication Date: 2013-04-03
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

At present, there is no duck hepatitis A virus type 3 vaccine in the market, so the prevention and treatment of duck hepatitis A virus type 3 is still blank in China

Method used

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  • DHAV (duck hepatitis A virus) JS strain and application of DHAV JS strain in duck virus hepatitis prevention and cure
  • DHAV (duck hepatitis A virus) JS strain and application of DHAV JS strain in duck virus hepatitis prevention and cure
  • DHAV (duck hepatitis A virus) JS strain and application of DHAV JS strain in duck virus hepatitis prevention and cure

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Embodiment 1: Isolation and cultivation of duck hepatitis A virus JS strain

[0036] 1 Disease material collection: Aseptically collect diseased materials such as liver and spleen of ducklings, cut the diseased materials into pieces, grind them finely, dilute them with sterile pH7. Antibiotics, 2000IU penicillin and 2000μg / mL streptomycin, act at 37°C for 30min, add chloroform to the centrifuged supernatant at a ratio of 4:1, mix well, place at 37°C for 60min, centrifuge at 3000rpm for 30min, take Antibiotics, 2000 IU penicillin and 2000 μg / mL streptomycin were added to the supernatant, and they were treated at 37°C for 30 min as materials for separating viruses.

[0037] 2 Isolation culture: inoculate 10-day-old SPF duck embryos with the above-mentioned virus isolation materials, 0.2mL / embryo, and incubate them in an incubator at 37°C to 38°C, illuminate the embryos 2 to 4 times a day, observe for 5 days, and incubate dead duck embryos after 48 to 120 hours. Place the...

Embodiment 2

[0038] Embodiment 2: the preparation of duck hepatitis A virus inactivated vaccine

[0039] 1. Establishment of basic seed batches: take the isolated freeze-dried virus seeds, dilute them with pH 7.4 PBS at 1:100, inoculate 10-day-old SPF duck embryos, 0.2mL / embryo, irradiate the embryos 2-4 times a day, observe for 5 days, and collect For duck embryos that died at 48 to 120 hours, the allantoic fluid and embryo bodies were preserved and tested for sterility, and fetal lesions were observed. Freeze and thaw repeatedly 3 times, centrifuge at 4000rpm for 30min, absorb the supernatant, aliquot and freeze-dry for storage.

[0040] 2. Establishment of production seed batches: take 12-day-old SPF duck embryos, inoculate basic virus seeds, incubate at 37-38°C, illuminate embryos 2-4 times a day, observe for 5 days, collect dead duck embryos at 48-120 hours, and take allantoic fluid And embryo body preservation and sterility test, and observation of fetal lesions. Freeze and thaw re...

Embodiment 3

[0050] Embodiment 3: the application of duck hepatitis A virus inactivated vaccine

[0051] 1. Immunization test of different doses and batches of inactivated duck hepatitis A virus vaccine on ducklings

[0052] Three batches of duck hepatitis A virus inactivated vaccines (prepared according to the method in Example 2) were inoculated with 1-day-old susceptible ducklings at 0.02mL / , 0.05mL / and 0.10mL / , and challenged duck hepatitis A 24h after inoculation Virus JS strain, 1000LD 50 / 0.2mL, observe the protection situation of ducklings, the results are shown in Table 1.

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Abstract

The invention discloses a novel DHAV (duck hepatitis A virus) JS strain and application of the DHAV JS strain in duck virus hepatitis prevention and cure. The DHAV JS strain is obtained through the steps of separation culture, duck embryo neutralization test, PCR (polymerase chain reaction) detection of viruses and sequencing detection experiment of the viruses, and the strain is determined to be the DHAV serotype 3; and the microbial preservation number is CGMCCNo.6852. According to the invention, through taking the novel DHAV JS strain as the virus strain for producing vaccine, the novel DHAV inactivated vaccine is prepared through the steps of inoculating 10-day-old SPF (specific pathogen free) duck embryos, selecting the embryos died after incubating for 48 hours, collecting the embryo liquid, inactivating and emulsifying. The clinical application proves that the prepared inactivated vaccine is favorable in safety and conducive to prevention and control of the novel DHAV disease, can prevent the infection and outbreak of the novel DHAV in a targeted way, and can be in mass production and clinical application.

Description

technical field [0001] The invention relates to an isolated virus strain and its application, in particular to a duck hepatitis A virus JS strain and its application in the prevention and treatment of duck viral hepatitis. It belongs to the field of biological vaccine prevention and control. Background technique [0002] Duck hepatitis virus (DHV) is the causative agent of duck viral hepatitis. The international virus classification system was revised in April 2010: the duck hepatitis virus was named duck hepatitis A virus (DHAV), which belongs to the picornaviridae family and the genus avian hepatitis virus. Currently duck hepatitis A virus is divided into three serotypes, namely: serotype 1, serotype 2 and serotype 3. Among them, serotype 1 is the traditional duck hepatitis virus, serotype 2 and serotype 3 are new virus strains discovered in recent years, corresponding to the new strains in Taiwan and South Korea, and there is no cross-reaction between the three serotype...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/125A61P31/14C12R1/93
Inventor 王彬藏玉婷邬立权柴华王琪宋扬刘洪斌涂映霞王峰张杨丁国杰
Owner 哈药集团生物疫苗有限公司
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