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Chicken new castle disease-infectious bronchitis trivalent combined vaccine and preparation method thereof

A technology for bronchitis and chicken Newcastle disease, which is applied in pharmaceutical formulations, medical preparations containing active ingredients, viral antigen components, etc., can solve the problems of cumbersome process, high cost, easy pollution, etc., and achieve the effect of overcoming side effects and improving the probability of prevention and treatment.

Inactive Publication Date: 2014-02-26
TIANJIN RINGPU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] There have been a large number of research reports on IBV and NDV dual vaccines in my country, but there are very few studies on the preparation methods of dual trivalent vaccines, and there are even fewer studies on renal type IBV and NDV dual trivalent vaccines. less
In 2006, Cui Baoan (patent application number: 200610018069.2) successfully prepared the IB / ND dual trivalent inactivated vaccine using the respiratory type M41 strain of IBV, the renal type HN99 strain and the La Sota strain of NDV, but it was inactivated by formaldehyde. Live-produced inactivated vaccines have the inherent shortcomings of traditional inactivated vaccines, and the immune effect is poor; in 2012, Song Xinyu (patent application number: 201210594755.X) used IBV glandular type YB160 and respiratory type H120 strain and La Sota strain of NDV prepared the IB / ND dual trivalent live vaccine, which gave full play to the advantages of the live vaccine and successfully attacked the glandular stomach type and respiratory type chicken infectious bronchitis virus currently prevalent in China The challenge with Newcastle disease virus provides good immune protection, but because chicken infectious bronchitis has the characteristics of poor cross-protection between different types of vaccines, so the product is still powerless to protect chicken kidney type IB
[0007] In the preparation process of traditional multiple vaccines, different viruses are often inoculated separately and expanded to prepare antigens. In the preparation process of the existing IB / ND dual trivalent vaccine, this antigen preparation mode is continued. It often has problems such as cumbersome process, high cost, easy pollution, and unstable potency between different batches.

Method used

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  • Chicken new castle disease-infectious bronchitis trivalent combined vaccine and preparation method thereof
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  • Chicken new castle disease-infectious bronchitis trivalent combined vaccine and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Example 1: Screening of Chicken Infectious Bronchitis Kidney Type K136 Strain

[0028] From 2003 to 2005, our company collected suspected chicken infectious bronchitis virus isolates from different chicken farms in my country. The sick chickens mainly manifested as lack of energy, coughing, and often heard grunting. Pathological autopsy found that there were obvious lesions in the kidneys in addition to small bleeding spots in the tracheal ring. After preliminary identification, 4 strains of renal-type infectious bronchitis virus were obtained. They were named CK / IBV / QD03 strain, CK / IBV / ZC03 strain, CK / IBV / YT04 strain and CK / IBV / SD05 strain respectively. Use 10-day-old SPF chicken embryos to carry out continuous subculture to attenuate the four strains. In each generation, 4 chicken embryos were inoculated through the allantoic cavity, incubated at 37°C, and the chicken embryos were frozen to death 36 hours after inoculation, and the allantoic fluid was harvested for th...

Embodiment 2

[0037] Embodiment 2: K136 strain safety detection

[0038] 1. Determination of toxicity of K136 strains of different generations on chicken embryos

[0039] Take the E2, E4, E7, E10, E15 generation virus seeds of the K136 strain, make appropriate multiple dilutions, inoculate 10-day-old SPF chicken embryos through the allantoic cavity, 0.1 mL per embryo, and set a blank control group at the same time, The cavity was inoculated with 0.1ml of physiological saline, and incubated at 37°C. The chicken embryos that died within 24 hours were discarded, and after 144 hours of incubation, the embryos were taken out to observe the development of the chicken embryos. Calculation of EID according to the Reed-Muench method 50 , the poisonous price of E2, E4, E7, E10, E15 generation poison species is respectively: 10 6.7 、10 6.8 、10 6.5 、10 6.6 、10 6.8 EID 50 / 0.1ml. The test results showed that the K136 strain proliferated stably on chicken embryos, and there was no reversion of v...

Embodiment 3

[0046] Example 3: Evaluation of immune efficacy of different generations of K136 strain

[0047] Use the E2, E4, E7, E10, E15 generation K136 strains to breed chicken embryo allantoic fluid in 5×10 2.0 EID 50 Dosage, intranasal drops for 1-day-old SPF chickens, 14 days after immunization, together with control chickens, each chicken used 10 3.68 EID 50 / 0.1ml of the isolated virulent CK / IBV / SD05 strain 0.05ml for eye drops and 0.05ml nasal drops respectively. On the 5th day after the challenge, the tracheal swabs of each chicken were collected and inoculated into the 10-day-old SPF chicken embryo 5 through the allantoic cavity. Embryo, 0.1ml / embryo, incubated for 144 hours. The results showed that the toxicity rate of the immune group was between 0 / 10-1 / 10, and the protection rate was between 9 / 10-10 / 10; the toxicity rate of the control group was 10 / 10. It shows that the immunogenicity of IBV-K136 strain virus seeds is basically the same within 15 generations, and they all...

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Abstract

The invention discloses a chicken new castle disease-infectious bronchitis trivalent combined vaccine and a preparation method thereof. The trivalent combined vaccine comprises antigens and a freeze-drying protective agent. The antigens comprise a chicken kidney-type infectious bronchitis virus K136 strain having an accession number of CCTCC-V201320, a chicken breath-type infectious bronchitis virus H120 strain, and a chicken new castle disease virus La Sota strain. A mode of inoculating the K136 strain and the La Sota strain in a homeomorphism manner and inoculating the H120 strain individually is adopted for preparing the antigens. Based on the mode, after culture products are mixed according to a ratio to prepare the antigens, a product of the vaccine is obtained by adding the freeze-drying protective agent. The trivalent combined vaccine can prevent the chicken new castle disease and the chicken infectious bronchitis simultaneously, thus solving effectively a problem that vaccines at present can only provide a part of protection or cannot provide protection for the kidney-type IB, and achieving an objective that one injection can be used for preventing two diseases.

Description

technical field [0001] The invention belongs to the technical field of new biomedicine for livestock and poultry, and in particular relates to a dual-combination trivalent live vaccine for chicken infectious bronchitis and chicken Newcastle disease and a preparation method thereof. Background technique [0002] Infectious bronchitis virus (IBV) belongs to the genus Coronaviridae of the Coronaviridae family and can cause an acute and highly contagious respiratory infectious disease in chickens. Its clinical symptoms mainly include dyspnea, rales, coughing, mouth breathing, Sneezing, etc., the mortality rate is generally very low, but chickens of various ages and sexes are susceptible to infection. Infection of young chickens often causes permanent degeneration of oviducts; infection of adult broilers usually results in reduced feed remuneration and increased mortality rate; infection of laying hens usually results in reduced egg production and lower egg quality. In addition,...

Claims

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Application Information

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IPC IPC(8): A61K39/295A61K39/17A61K9/19A61P31/14A61K39/215
Inventor 陈冰何召庆杨保收孙美玉邢婧珉李云霞梁武李守军
Owner TIANJIN RINGPU BIO TECH
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