Hog cholera virus genetic recombination adenovirus carrier vaccine rabbit body effect detection method

A technology of gene recombination and swine fever virus, applied in the field of biotechnology detection, can solve the problems of no stable, simple and cost-saving vaccine efficacy evaluation standards, etc.

Inactive Publication Date: 2015-09-30
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

rAd-E0-E2 is the latest genetically engineered live vector vaccine developed by our laboratory, since it does not have a more stable, simple and cost-effective vaccine efficacy evaluation standard

Method used

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  • Hog cholera virus genetic recombination adenovirus carrier vaccine rabbit body effect detection method
  • Hog cholera virus genetic recombination adenovirus carrier vaccine rabbit body effect detection method
  • Hog cholera virus genetic recombination adenovirus carrier vaccine rabbit body effect detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Establishment of the Efficacy Testing Method for Rabbit Body of New Type CSFV Gene Recombinant Adenovirus Vector Vaccine

[0018] 1.1 Experimental poisons, vaccines, and experimental animals

[0019] Recombinant adenovirus vaccine for classical swine fever (rAd-E0-E2, ≥2×10 7.0 IFU / bottle, the determination method is detailed in the patent: Zhang Heng, Fan Gencheng, Du Yuanzhao, etc. A method for determining the virus content of the recombinant adenovirus vector vaccine of classical swine fever virus [P]. The State Intellectual Property Office of the People's Republic of China. CN 104535764 A, 2015.04.22), was freeze-dried and preserved by the technology center of Qingdao Yibang Bioengineering Co., Ltd.; live swine fever vaccine (rabbit source) referred to as swine fever spleen leaching virus, batch number 201401, 20 heads / bottle, produced by the freeze-dried seedling workshop of the company ; 1.5-3kg healthy susceptible rabbits were purchased from Qingdao K...

Embodiment 2

[0035] Embodiment 2: Rabbit body potency testing method determines the minimum immune dose of rAd-E0-E2

[0036] The minimum immune dose of rAd-E0-E2 is determined according to the rabbit body efficacy test method established in Example 1, and the steps are as follows:

[0037] 2.1 Immunization method Carry out immunization according to the method in Table 1 in step 1.2.

[0038] 2.2 Immunization methods and actual temperature measurement results before and after immunization The body temperature was measured in the morning and afternoon for 3 consecutive days before and after immunization.

[0039] 2.3 Protective effect of rAd-E0-E2 on immunized rabbits

[0040] After attacking the poison, the original multiplier (10 7.0 IFU) immunized rabbits and CSF splenic gonorrhea immunized rabbits had body temperature 6.0 3 / 4 of IFU) immunized rabbits had a stereotyped thermal reaction, 3 / 4 of the rabbit spleen / body mass index ratio was relatively large, 3 / 4 of the spleen tissue secti...

Embodiment 3

[0045] Embodiment 3: Rabbit physical effect test method evaluates the effectiveness of 3 batches of freeze-dried rAd-E0-E2 vaccines

[0046] Get 3 batches of freeze-dried rAd-E0-E2 vaccine (potency ≥ 2 × 10 7.0 IFU / bottle), named as freeze-dried vaccine 1, freeze-dried vaccine 2 and freeze-dried vaccine 3 respectively. The 3 batches of vaccines were immunized according to the method in Table 1 in 1.2, and the five evaluation methods of the effect test model established in the specific example 1 were used for comparison and detection. The results are shown in Tables 3-5. According to the comparative test results, it was proved that the potency of the 3 batches of freeze-dried vaccines was relatively stable, and it was further determined that the minimum immune dose of the vaccines was all at 10 7.0 IFU protection, and further prove the stability of the rabbit physical examination method.

[0047] Table 3 Comparison of protection results of five evaluation methods after immuni...

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Abstract

The invention relates to establish a detection method mode for hog cholera virus genetic recombination adenovirus carrier vaccine rabbit body effect. An SPF rabbit which is stable in production performance and quality serves as a carrier, after rAd-Eo-E2 immunization is subjected to a rabbit body for 14 days, swine fever spleen vaccine serves as strain used for counteracting toxic substances, after the toxic substances is counteracted, rabbit body heat of reaction is monitored continuously for 3 days, the protective efficacy of the vaccine is evaluated according to situation of forming heat (>= 40.5 DEG C) occurring on the rabbit body, and the detection method mode for the for rAd-Eo-E2 rabbit body efficacy is established. In the method establishing process, by means of temperature monitoring, an index ratio of the spleen to weight of the rabbit, spleen tissue section observation, RT-PCR detection and swine fever spleen gonorrhea separation and identification of an immune group and a matched group, whether the protective efficacy of the rAd-Eo-E2 immunization displayed by the result is accordant or not is determined, and evaluation conducted by adopting a relatively simple, convenient and easy detection method of temperature monitoring is determined finally.

Description

technical field [0001] The invention belongs to the field of biotechnology detection, and in particular relates to a method for testing the effectiveness of a novel swine fever virus gene recombinant adenovirus vector vaccine. Background technique [0002] Classical swine fever (CSF) is a highly contagious infectious disease of pigs caused by Classical swine fever virus (CSFV) of the genus Pestiviridae in the Flaviviridae family. Economic losses. In the face of such severe infectious diseases, our country generally uses vaccines and vaccinations to prevent them in advance to reduce morbidity and mortality. The development of swine fever vaccine has experienced early inactivated vaccines, attenuated vaccines, and genetically engineered vaccines. Attenuated vaccines are widely used at present. The attenuated vaccine approved for use in my country is the attenuated swine fever rabbit vaccine developed by the China Veterinary Drug Administration. For a long time, the widespre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00C12Q1/70C12Q1/68G01N21/00
Inventor 张恒张小苗范根成杜元钊孙永科孙成友申洪银胡潇郭玉广陶晓珊王龙
Owner YEBIO BIOENG OF QINGDAO
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