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An Isolate Strain of Infectious Bronchitis Virus and Its Application in Vaccine Preparation

A virus isolation and bronchitis technology, applied in the direction of virus antigen components, viruses, vaccines, etc., can solve the problems of strain variation, increased prevention and control of infectious bronchitis, difficulty, etc., and achieve significant immune efficacy.

Active Publication Date: 2021-08-13
BEIJING HUAXIA XINGYANG BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the current IBV mutation, the number of serotypes has increased, and the existing vaccine strains can only provide protection for the corresponding serotypes. However, if the attenuated vaccine for infectious bronchitis is used and operated improperly, or after several years of continuous use, there will be certain risk of infection. Pathogenicity and as the main body of mutations or donors of recombinant mutations, resulting in strain mutations, making the prevention and treatment of infectious bronchitis more difficult

Method used

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  • An Isolate Strain of Infectious Bronchitis Virus and Its Application in Vaccine Preparation
  • An Isolate Strain of Infectious Bronchitis Virus and Its Application in Vaccine Preparation
  • An Isolate Strain of Infectious Bronchitis Virus and Its Application in Vaccine Preparation

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Experimental program
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Effect test

Embodiment 1

[0018] Embodiment 1 Isolation and identification of chicken infectious bronchitis virus cb2 strain of the present invention

[0019] 1.1 Virus isolation

[0020] A case of suspected infectious bronchitis symptoms appeared in a farm in Shandong. Autopsy of dead chickens showed mucus in the trachea, blockage of the trachea, and obvious "spotted kidneys". Preliminary diagnosis was that infectious bronchitis virus caused morbidity and death. Collect the trachea, kidneys and lungs of sick and dead chickens, mix them, cut them into pieces, grind them, dilute them with sterile PBS according to the W / V ratio of 1:5, freeze and thaw them three times, centrifuge them at 3800rpm for 10min, and take the supernatant and filter them with a 0.22μm microporous filter. Membrane filtration, the filtrate was used to inoculate five 10-day-old SPF chicken embryos through the allantoic cavity at a dose of 0.2ml / embryo. After inoculation, the chicken embryos were placed in a 37°C incubator to cont...

Embodiment 2

[0033] Example 2 Comparison of Infectious Bronchitis Virus S1 Gene RT-PCR, Sequencing and Evolutionary Analysis

[0034] Virus RNA was extracted from the allantoic fluid of the 4th generation chicken embryo of the cb2 strain according to the instructions of TRIzol. The RT-PCR method was used to amplify the virus S1 gene. The recovered PCR products were sent to Shanghai Sangon Bioengineering Technology Service Co., Ltd. for sequencing. Analyze the sequence of the virus S1 gene and compare it with the reference strains in GenBank, see attached figure 1 : Evolutionary relationship of gene sequence of virus spike protein S1 subunit of vaccine strain cb2 strain.

[0035] The newly isolated cb2 strain of infectious bronchitis is closely related to most domestic isolates (such as QX strains and QX-like strains), so it may form cross-protection against current epidemic strains. The new isolate infectious bronchitis cb2 is more distantly related to the S1 gene of the currently used ...

Embodiment 3

[0036] Example 3 Safety and effectiveness test of chicken infectious bronchitis inactivated vaccine cb2 strain of the present invention

[0037] 3.1 Preparation and inspection of antigens for seedling production

[0038] Take the virus seed for production, dilute it 1000 times with sterilized normal saline, inoculate 11-day-old non-immune chicken embryos, inoculate 0.2ml into the allantoic cavity of each embryo, seal the pinhole after inoculation, and continue to incubate at 36-37°C. Turn the eggs. After inoculation, the eggs were illuminated twice a day, and the chicken embryos that died within 30 hours were discarded. Take out the chicken embryos that died within 30-48 hours at any time, and after 48 hours of cultivation, regardless of whether they died or not, all of them were taken out, with the air chamber upward, and placed at 2-8°C to cool for 24 hours. Take out the cooled chicken embryo, disinfect the egg shell of the air cell, and then use aseptic surgery to peel of...

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Abstract

The invention relates to an isolated strain of infectious bronchitis virus and its application in vaccine preparation. The strain was named cb2 strain, and it was preserved in the General Microbiology Center of China Microbiological Culture Collection Management Committee on June 21, 2016, and the microbial preservation number is CGMCC No.12657. The invention adapts the isolated virulent strain of infectious bronchitis virus to chicken embryos, and obtains a virus strain with high titer suitable for the growth of chicken embryos. A stable pathogenesis model can be established by using the virus strain, and the virus strain maintains good specificity and immunogenicity, and can be used for the production and preparation of vaccines. After the virus strain of the present invention is inactivated, the inactivated vaccine is prepared into an inactivated vaccine for immune efficacy and safety evaluation tests. The results show that the vaccine is safe to the target animals, has no local and systemic adverse reactions caused by the vaccine, and can effectively prevent kidney-type and respiratory-type infections. Infection caused by bronchitis virus.

Description

technical field [0001] The invention belongs to the field of preventive veterinary veterinary biological products, in particular relates to chicken infectious bronchitis virus strains and vaccine compositions thereof, in particular to the application of chicken infectious bronchitis virus strains in the preparation of inactivated vaccines. Background technique [0002] Chicken infectious bronchitis (Avian Infectious Bronchitis, IB) is an acute, highly contagious, viral respiratory disease caused by chicken infectious bronchitis virus (Infectious Bronchitis Virus, IBV). virus genus. Chickens infected with infectious bronchitis virus: cough, tracheal rales; kidney enlargement, paleness, ureter dilatation, a large amount of urate deposition, and the appearance of a typical "mottled kidney". The virus is not resistant to the environment, is allergic to common disinfectants, and has a certain resistance to low temperature. [0003] Infectious bronchitis virus has strong variabi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/215A61P31/14C12R1/93
CPCA61K39/12A61K2039/5252A61K2039/552A61P31/14C12N7/00C12N2770/20021C12N2770/20034
Inventor 赵卓尚川川胡义彬贺亚奇商云鹏吴峰郭小燕张爽李晓亮王顺山金扩世王力江厚生
Owner BEIJING HUAXIA XINGYANG BIOLOGICAL SCI & TECH
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