Antibody testing method for efficacy of a classical swine fever genetically engineered subunit vaccine

A subunit vaccine and genetic engineering technology, applied in the field of veterinary biological products, can solve the problems of long testing period, shortening testing period, high cost of feeding and testing, and achieve short testing period, saving testing cost and high biological safety. Effect

Active Publication Date: 2018-11-20
YEBIO BIOENG OF QINGDAO
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The genetically engineered subunit vaccine of CSFV is an inactivated vaccine. Except for the efficacy test method of using the target animal (pig) for immune challenge, there is currently no alternative method available. The detection period of the method is long (51 days), the cost of breeding and inspection is high, and the inspection process involves the use of virulence of classical swine fever. In order to solve the above problems, it is urgent to establish a feasible and effective experimental animal efficacy inspection method for the efficacy inspection of inactivated vaccines. It can shorten the detection cycle, save detection cost and improve biological safety

Method used

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  • Antibody testing method for efficacy of a classical swine fever genetically engineered subunit vaccine
  • Antibody testing method for efficacy of a classical swine fever genetically engineered subunit vaccine
  • Antibody testing method for efficacy of a classical swine fever genetically engineered subunit vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1: A kind of swine fever genetic engineering subunit vaccine (293T-E2) potency testing method

[0019] Include the following steps:

[0020] (1) Get 20 healthy susceptible rabbits (rabbits whose ELISA antibody is negative for classical swine fever virus) as test objects, and randomly divide them into 4 groups according to body weight and sex, with 5 rabbits in each group; 3 groups are immune groups, 1 group is the control group;

[0021] (2) Inject the rabbits in the immunized group. Three groups are injected with 0.1ml, 0.2ml, and 0.5ml of the CSF genetically engineered subunit vaccine (293T-E2) respectively, while the rabbits in the control group do not inject any substance; inject the vaccine The rabbits in the control group and the rabbits in the control group were raised according to the normal feeding method;

[0022] (3) After 21 days of step (2) injection of the vaccine, the rabbits in the immunization group and the control group were all subjected ...

Embodiment 2

[0027] Example 2: Correlation detection between antibody detection and virus attack protection:

[0028] In embodiment 1, after the rabbits of the immunization group and the control group carried out heart blood collection, each rabbit carried out ear vein injection of live vaccine for swine fever (spleen leaching source), 1.0ml / only; The body temperature was measured once a day for the first two days of the injection, and every 6 hours after the injection, for a total of 72 hours; when the temperature change of the rabbit before and after the injection did not exceed 1°C, it was determined that the protective effect had been achieved. The results are shown in Table 2:

[0029] Table 2: Attack Protection

[0030] group

[0031] It can be seen from Table 2 that after the rabbits are injected with the CSF genetically engineered subunit vaccine (293T-E2), they are then challenged with the CSF virus (the attenuated strain of CSF in the CSF splenogenous live vaccine), wh...

Embodiment 3

[0032] Embodiment 3: Application of the test method of the present invention and comparison with target animal test

[0033] Take 20 healthy and susceptible rabbits weighing 1.5-3.0kg (the ELISA antibodies of pestivirus are all negative) and divide them into 4 groups at random, with 5 rabbits in each group, and inject 201401, 201402, and 201403 batches of pigs subcutaneously in each neck of groups 1-3 Pestilence gene engineering subunit vaccine (293T-E2), 0.2ml / only, the 4th group is the control group. On the 21st day after immunization, blood was collected from the heart of all rabbits, centrifuged at 7000r / min for 10 minutes, and the supernatant was drawn to detect the ELSIA antibody to classical swine fever; if the antibody was positive, it proved that the genetic engineering subunit vaccine (293T-E2) had achieved protective effect. The results are shown in Table 3:

[0034] Table 3: Antibody detection

[0035]

[0036] Note: Blocking rate ≥ 40% is positive (+); 30% < ...

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Abstract

The invention provides a method for testing potency of a CFS (classical swine fever) virus genetic engineering subunit vaccine. The method comprises steps as follows: healthy susceptible rabbits are taken as test objects and divided into a control group and immunization groups; the CFS virus genetic engineering subunit vaccine is injected into rabbits of the immunization groups while no substances are injected to rabbits of the control group; after 21 days of vaccine injection, the rabbits of the immunization groups and the control group are subjected to blood sampling from hearts, heart blood is centrifuged, then liquid supernatant is taken, a CFS ELSIA (enzyme linked immunosorbent assay) antibody is detected, and when at least 4 / 5 of the antibody is positive, the test for the CFS virus genetic engineering subunit vaccine is qualified. The method for testing the potency of the CFS virus genetic engineering subunit vaccine can replace a potency testing method adopting swine for immunity challenge; the test animals, namely, the rabbits, are easily available, so that the testing cost is reduced, the testing period is short, CFS live virus infection is not involved, and biological safety is high.

Description

technical field [0001] The invention relates to an antibody test method for the effectiveness of a classical swine fever genetic engineering subunit vaccine (293T-E2), belonging to the field of veterinary biological products. Background technique [0002] Swine fever (slassical swine fever, CFS) is characterized by hemorrhage and fever, and can be acute or chronic. In many countries and regions in the world, traditional vaccination is an important means of controlling swine fever. The currently used attenuated vaccines are mainly Chinese swine fever rabbitized attenuated vaccine, Japanese GPE vaccine and French Thireval vaccine. In the late 1970s, the epidemic form of swine fever changed a lot. Regional sporadic epidemics and the occurrence of atypical swine fever symptoms, especially in recent years when monoclonal antibodies have been used at home and abroad to detect C-strains, different reaction patterns have been found, and swine fever immunization has repeatedly fail...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983G01N33/94G01N2333/08
Inventor 刘蕾张恒陶晓珊邹敏孙健王丽萍赵同渊
Owner YEBIO BIOENG OF QINGDAO
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