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Chicken infectious bursal disease virus classical strain for natural adaptation to in vitro cell culture and application of chicken infectious bursal disease virus classical strain

A technology for chicken infectivity and bursal disease, which is applied in the direction of viruses, antiviral agents, and viral antigen components, can solve problems such as poor antigen matching, high R&D costs, and antigenic variation of virus strains, and achieve the effect of solving bottlenecks

Active Publication Date: 2020-06-05
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, wild IBDV strains that can adapt to in vitro cell culture such as chicken embryo fibroblasts (CEF cells) and their subcultured cell line DF1 have not been found in nature, so the current IBDV whole virus vaccine strains on the market are all derived from artificial subculture
The artificial passage technology of infectious bursal disease virus (IBDV) has the following disadvantages: ① The process is time-consuming and laborious, with high research and development costs and poor timeliness
②During the passaging process, the antigenicity of the virus strain may mutate, resulting in poor antigen matching between the vaccine strain and the epidemic strain
③ The subculture results are random, and not every subculture test can obtain the ideal strain

Method used

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  • Chicken infectious bursal disease virus classical strain for natural adaptation to in vitro cell culture and application of chicken infectious bursal disease virus classical strain
  • Chicken infectious bursal disease virus classical strain for natural adaptation to in vitro cell culture and application of chicken infectious bursal disease virus classical strain
  • Chicken infectious bursal disease virus classical strain for natural adaptation to in vitro cell culture and application of chicken infectious bursal disease virus classical strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1I

[0035] Isolation and Identification of Example 1 IBD17JL01 Strain

[0036] 1. Test materials and methods

[0037] 1.1 Collection and processing of clinical samples

[0038] In September 2017, 25-day-old laying hens on a laying hen farm in Fuyu City, Jilin Province had clinical symptoms suspected of IBD. The sick chicken showed lack of energy, and the necropsy found that its bursa was severely atrophied, and hemorrhage points could be seen in the leg muscles and bursa. Add an appropriate amount of PBS to the sick chicken bursa sample, grind and freeze-thaw three times, and centrifuge the prepared suspension at 5000g for 5min at 4°C, and take the supernatant for subsequent detection.

[0039] 1.2 Main reagents and materials

[0040] Prime STARTM HS DNA Polymerase, pMD18-T vector, RNAiso Plus, and DL2000DNAMarker were purchased from Dalian Takara Bioengineering Co., Ltd. (Takara); nucleic acid gel recovery kit and small plasmid kit were all products of AxyPrep; DMEM medium was...

Embodiment 2I

[0072] The immune efficacy test of embodiment 2IBD17JL01 strain inactivated vaccine

[0073] 1. Materials and Methods

[0074] 1.1 Main reagents and instruments

[0075] The white oil adjuvant was purchased from Harbin Veken Biotechnology Co., Ltd.; the IBDV antibody ELISA detection kit was purchased from IDEXX Company of the United States; the multifunctional microplate reader was a product of PE Company of the United States.

[0076] 1.2 Test animals

[0077] SPF chickens were purchased from the Experimental Animal Center of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences; SPF chickens were kept in the negative pressure isolator of the center.

[0078] 1.2 Viruses

[0079] IBDV classic strain IBD17JL01 strain (preservation number is CGMCC No.19291) was isolated and identified in Example 1. The IBD17JL01 strain can be adapted to in vitro cell culture such as DF1. In this study, the IBD17JL01 strain was amplified on DF1 cells, and the seed b...

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Abstract

The invention discloses a chicken infectious bursal disease virus classical strain for natural adaptation to in vitro cell culture and an application of the chicken infectious bursal disease virus classical strain. The chicken infectious bursal disease virus classical strain is named as IBD17JL01, and is preserved in CGMCC(China General Microbiological Culture Collection Center), wherein the preservation No. is CGMCC No.19291. The virus strain is the chicken infectious bursal disease virus classical strain for natural adaptation to DF1 and CEF in vitro cell culture, which is isolated and identified for the first time. The virus strain can directly proliferate on DF1 cells, CEF cells or DT40 cells of an in vitro passage cell line. After being inactivated, the virus strain is prepared into an inactivated vaccine, so that immunised chicks can generate 100% of protection effects on virus attack. Therefore, the virus strain ( IBD17JL01 strain) can be used as a vaccine candidate strain for natural adaptation to in vitro cell culture to be used for immunization control of IBDV.

Description

technical field [0001] The invention relates to a novel chicken infectious bursal disease virus strain and its application, in particular to a chicken infectious bursal disease virus classic strain naturally adapted to in vitro cell culture and its application. The invention belongs to the field of biotechnology. Background technique [0002] Infectious bursal disease (IBD) is an important immunosuppressive disease of chickens, and its pathogen is infectious bursal disease virus (IBDV). IBDV has two serotypes: serotype I and serotype II. Serum type I is pathogenic to chickens, while serum type II is not pathogenic to chickens. Serum type I is divided into classic strains, super-virulent strains and mutant strains, and the antigenicity of classic strains and super-virulent strains is the same. Infectious bursal disease caused by classic strains of IBDV first occurred in the United States in 1957. In the late 1980s, IBDV variants and super-virulent strains appeared one aft...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/12A61P31/14C12R1/93
CPCA61K39/12A61K2039/5252A61K2039/552A61P31/14C12N7/00C12N2720/10021C12N2720/10034C12N2720/10051
Inventor 祁小乐王笑梅高玉龙高立李凯崔红玉刘长军潘青张艳萍
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI