Porcine pseudorabies virus positive serum preparation method

A porcine pseudorabies virus-positive serum technology is applied in the field of preparation of porcine pseudorabies virus-positive serum, which can solve problems such as increased social instability factors, biological safety hazards, and difficulty in ensuring purity, and reduce the risk of exogenous virus contamination , high school and valence effects

Active Publication Date: 2016-07-06
兆丰华生物科技(南京)有限公司 +1
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  • Summary
  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, many vaccines in my country do have biological safety hazards, which have caused major economic losses to farmers, people panic, and social instability factors have increased. Therefore, it is necessary to continuously improve and increase inspection and operation items during the production process to prevent contamination of exogenous viruses. leading to immune failure or triggering epidemic outbreaks caused by other viruses
[0003] At present, there are major constraints on exogenous virus testing for porcine pseudorabies virus live vaccines, that is, there is no high-potency commercialized low-cost positive serum, which has a certain impact on product testing
At present, although pigs are also used to prepare positive serum for porcine pseudorabies virus, the titer is not high, and can only reach 1:512, and because of the use of homologous animals, it may be more complicated to carry exogenous antigens and antibodies, and the purity It is difficult to guarantee, and it is also difficult to apply in the actual inspection process

Method used

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preparation example Construction

[0024] A preparation method for porcine pseudorabies virus positive serum, comprising the following steps:

[0025] First, prepare the inactivated vaccine:

[0026] 1) Preparation of transfer factor adjuvant: mix the organic phase with the transfer factor solution at a volume ratio of 1:1 to obtain the transfer factor adjuvant, the organic phase is a mixture of lecithin, cholesterol and Tween-80, the organic phase The molar ratio of lecithin and cholesterol in the phase is 1:1, and the concentration of the transfer factor solution is 6-8mg / ml;

[0027] 2) Preparation of the oil phase: mix white oil, Tween-80 and aluminum stearate according to the volume ratio of 94:6-7:1.5-2 to obtain the oil phase;

[0028] 3) Preparation of the water phase: in the porcine pseudorabies virus antigen solution (titer is 10 7.5 TCID 50 Add porcine pseudorabies virus antigen solution 1-1.5‰ formaldehyde to get water phase, then place the water phase in 37℃ water bath for 21-24h for inactivatio...

Embodiment 1

[0038] A preparation method for porcine pseudorabies virus positive serum, comprising the following steps:

[0039] First, prepare the inactivated vaccine:

[0040] 1) Preparation of transfer factor adjuvant: mix the organic phase with the transfer factor solution at a volume ratio of 1:1 to obtain the transfer factor adjuvant, the organic phase is a mixture of lecithin, cholesterol and Tween-80, the organic phase The molar ratio of lecithin and cholesterol in the phase is 1:1, and the concentration of the transfer factor solution is 6mg / ml;

[0041] 2) Preparation of the oil phase: white oil, Tween-80 and aluminum stearate were mixed according to the volume ratio of 94:6:1.5 to obtain the oil phase;

[0042] 3) Preparation of the water phase: in the porcine pseudorabies virus antigen solution (titer is 10 7.5 TCID 50Add 1‰ formaldehyde of porcine pseudorabies virus antigen solution per ml) to obtain the water phase, then place the water phase in a 37°C water bath for 21 ho...

Embodiment 2

[0052] A preparation method for porcine pseudorabies virus positive serum, comprising the following steps:

[0053] First, prepare the inactivated vaccine:

[0054] 1) Preparation of transfer factor adjuvant: mix the organic phase with the transfer factor solution at a volume ratio of 1:1 to obtain the transfer factor adjuvant, the organic phase is a mixture of lecithin, cholesterol and Tween-80, the organic phase The molar ratio of lecithin and cholesterol in the phase is 1:1, and the concentration of the transfer factor solution is 8mg / ml;

[0055] 2) Preparation of the oil phase: white oil, Tween-80 and aluminum stearate were mixed according to the volume ratio of 94:7:2 to obtain the oil phase;

[0056] 3) Preparation of the water phase: in the porcine pseudorabies virus antigen solution (titer is 10 7.5 TCID 50 / ml) by adding 1.5‰ formaldehyde in porcine pseudorabies virus antigen solution to obtain the water phase, and then place the water phase in a 37°C water bath f...

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Abstract

The invention discloses a porcine pseudorabies virus positive serum preparation method. A water-in-oil type inactivated vaccine is prepared through emulsification, and then porcine pseudorabies virus positive serum with a high neutralizing titer is obtained through immune reaction generated by stimulating sheep through the synergistic effect of a porcine pseudorabies virus live vaccine and the inactivated vaccine. The method reduces the risk of exogenous virus contamination caused by homologous animals when positive serum is prepared from pigs, and a new thought is provide for preparation of positive serum.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for preparing porcine pseudorabies virus positive serum. Background technique [0002] With the development of the times and the country, biosafety has been paid more and more attention by the country and the people, and the biosafety of biological products produced with viruses or bacteria as raw materials has gradually become the top priority of government supervision. At the same time, many vaccines in my country do have biological safety hazards, which have caused major economic losses to farmers, people panic, and social instability factors have increased. Therefore, it is necessary to continuously improve and increase inspection and operation items during the production process to prevent contamination of exogenous viruses. And lead to immune failure or trigger epidemic outbreaks caused by other viruses. [0003] At present, there are relatively large restrictive facto...

Claims

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Application Information

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IPC IPC(8): C07K16/08C07K16/06
CPCC07K16/06C07K16/085
Inventor 王玉玲赖道华黄玉婷詹先强
Owner 兆丰华生物科技(南京)有限公司
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