Method for testing efficacy of porcine Seneca virus disease inactivated vaccine by using rabbits

A technology of inactivated vaccines and viral diseases, applied in veterinary vaccines, vaccines, viruses, etc., can solve the problems of no longer safe and reliable, rising prices, and difficulties in purchasing experimental pigs, and achieve simple operation and low feeding costs , the effect of low purchase cost

Active Publication Date: 2020-10-02
JINYUBAOLING BIO PHARMA CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The prevention and treatment of the disease is mainly based on prevention. However, there is no commercialized vaccine for Seneca virus disease in my country at present, and the experimental vaccine is in the stage of research and development.
Among them, the efficacy test of the Seneca virus test vaccine mainly relies on pigs. Due to the impact of African swine fever, the trading and transportation of pigs are restricted, and the price has risen. Moreover, the procurement of experimental pigs is very difficult and is no longer safe and reliable, which directly leads to Research and development of inactivated vaccine against Cavirus disease hindered

Method used

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  • Method for testing efficacy of porcine Seneca virus disease inactivated vaccine by using rabbits
  • Method for testing efficacy of porcine Seneca virus disease inactivated vaccine by using rabbits
  • Method for testing efficacy of porcine Seneca virus disease inactivated vaccine by using rabbits

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1, the preparation of porcine Seneca virus inactivated vaccine

[0025] This embodiment chooses to use SVV / CH / ZZ / 2016CGMCC No.14886 (CN110551694A) as the strain to prepare Seneca virus liquid, obtain the vaccine in the following examples after inactivation and emulsification, specifically comprising the following steps:

[0026] Inoculate the Seneca virus strain at a cell density of 3.0 × 10 6 , PK15-JY suspension cells with cell viability above 95% (porcine kidney suspension cells, provided by Jinyu Baoling Biopharmaceutical Co., Ltd.), placed in 5% CO at 37°C 2 After culturing in the incubator for 3 days, the virus culture medium was harvested by repeated freezing and thawing three times, and BEI (diethyleneimine, Sigma) was added to make the final concentration 0.003mol / L, and inactivated at 26°C for 48 hours. According to the content of 4% (V / V), 50% sodium thiosulfate solution was added to block the inactivation, and the inactivated antigen was obtained...

Embodiment 2

[0027] Embodiment 2, the preparation of Seneca virus liquid

[0028] This embodiment selects and uses SVV / CH / ZZ / 2016 CGMCC No.14886 (CN110551694A) to prepare Seneca virus liquid as virus strain, is used for the challenge poison in following embodiment, specifically comprises the following steps:

[0029] The Seneca virus strain was inoculated on PK-15 cells (porcine kidney cells, provided by Jinyu Baoling Biopharmaceutical Co., Ltd.) overgrown with a single layer, and placed in 5% CO at 37°C. 2 Cultivate in the incubator for 5 days, freeze and thaw repeatedly 3 times to harvest the virus culture medium, record it as the F1 generation, and store it at -20°C. Take the F1 generation of the harvested virus culture medium, inoculate PK-15 cells that have grown into a monolayer according to the content of 10% (V / V), and place them in 5% CO at 37°C. 2 Cultivate in an incubator, observe cytopathic changes (CPE) every day, harvest the virus culture liquid 48 hours after inoculation, a...

Embodiment 3

[0030] Embodiment 3, determination of rabbit challenge dose

[0031] This embodiment utilizes the Seneca virus liquid that above-mentioned embodiment 2 obtains to carry out challenge test to rabbit, to determine the challenge dose of rabbit, specifically comprises the following steps:

[0032] 3.1. Selection of experimental animals: 20 healthy and susceptible rabbits of 1.0-2.0 kg were selected, and the serum neutralizing antibody of Seneca virus was detected <1:2, purchased from Eternal Agriculture and Animal Husbandry Development Co., Ltd., Dalat Banner, Ordos City;

[0033] 3.2. Preparation of poison for attacking poison: the poison price prepared in Example 2 is 10 7.5 TCID 50 / 0.1ml of the virus liquid is the virus stock solution for gradient dilution to obtain a 10-fold dilution, 10 2 double dilution and 10 3 Two-fold dilutions are used to challenge the rabbits respectively;

[0034] 3.3. Virus challenge: 20 healthy susceptible rabbits were randomly divided into 5 gr...

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Abstract

The invention discloses a method for testing the efficacy of a porcine Seneca virus disease inactivated vaccine by using rabbits, and belongs to the field of veterinary biological products. The methodcomprises the following steps: performing immunization on a plurality of healthy susceptible rabbits by using the porcine Seneca virus disease inactivated vaccine; after 28 days of immunization, performing virus attack on the immunized rabbits by using a Seneca virus solution; and after virus attack, continuously observing for 15 days, and when at least 4 / 5 of the rabbits do not show clinical symptoms after virus attack, judging that the protection efficacy of the porcine Seneca virus disease inactivated vaccine is qualified. According to the method, the rabbits can be effectively used for replacing experimental pigs to establish a replacement method for testing the efficacy of the Seneca virus disease inactivated vaccine, so that the cost is effectively reduced; and the method is simpleand feasible to operate, safe and convenient.

Description

technical field [0001] The invention belongs to the field of veterinary biological products, in particular to a method for testing the effectiveness of porcine Seneca virus disease inactivated vaccines with rabbits, and in particular to a porcine Seneca virus disease inactivated vaccine that can replace pigs for immune challenge tests Methods for testing the potency of live vaccines. Background technique [0002] Seneca virus (Seneca virus A, SVA) is a virus that can cause vesicle-like symptoms in pigs. The virus is an RNA virus with a single-stranded positive strand and no segmentation, belonging to the Senecavirus genus of the small RNA virus family. Clinical evidence shows that after the virus infects pigs, it can cause vesicular lesions in infected pigs and death of newborn piglets. The symptoms are very similar to foot-and-mouth disease. , which is prone to confusion. The virus was first discovered in 1998, and gradually gained attention after a large-scale outbreak o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00A61K39/125A61P31/14A01K67/02
CPCA61K49/0008A61K39/12A61P31/14A01K67/02A61K2039/5252A61K2039/552A61K2039/545A61K2039/54C12N2770/32034Y02A50/30
Inventor 齐志涛陈九连路荣方建国高艳华王娥娥张云星郝苏云张燕红杜宇荣宋庆庆赵丽霞李雪峰毕力格
Owner JINYUBAOLING BIO PHARMA CO LTD
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