Preparation method of complex live vaccine for porcine reproductive and respiratory syndrome

A technology of respiratory syndrome and compound live vaccine, which is applied in the field of preparation of porcine reproductive and respiratory syndrome compound live vaccine, can solve the problems of high production cost, expensive vaccine, time-consuming and labor-intensive, etc., to ensure stability and shorten the cultivation time , reduce the effect of side effects

Active Publication Date: 2014-01-08
浙江美保龙生物技术有限公司
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  • Summary
  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The traditional antigen-antibody complex vaccine production method is to combine hyperimmune serum with bursal virus. This method requires a large amount of hyperimmune serum antibodies to be obtained by slaughtering a large number of animals, which is time-consuming and laborious. At the same time, the production cost is high, resulting in Vaccines are more expensive

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  • Preparation method of complex live vaccine for porcine reproductive and respiratory syndrome
  • Preparation method of complex live vaccine for porcine reproductive and respiratory syndrome

Examples

Experimental program
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Embodiment 1

[0049] Example 1 Production of porcine reproductive and respiratory syndrome virus (CH-1R strain) with torrent perfusion reactor AP20C type, comprising the steps:

[0050] (1) Selection of bioreactor: Rapid flow reactor type AP-20C with a volume of 10L.

[0051] (2) Selection of vaccine strains: Porcine Reproductive and Respiratory Syndrome Virus CH-1R strain was selected, the microorganism preservation number: CGMCC NO.1883, the preservation time was December 11, 2006, and the preservation unit was China Microorganism Culture Collection Management Committee General Microbiology Center for existing strains.

[0052] (3) Cultivation of different cells: Propagate Marc-145 or MA104 cells in a T175 square bottle, generally passaging at a ratio of 1:3 to 1:5, digest and count the cells after the cells have grown into a single layer, and inoculate the cell suspension into a transfer tube Inoculate about 3×10 cells per tube 7 cells were cultured in a microreactor, the temperature o...

Embodiment 2

[0060] The preparation of embodiment 2 traditional Chinese medicine polysaccharide comprises the following steps:

[0061] a. 10-30 parts of raw land, 20-40 parts of Achyranthes bidentata, 10-20 parts of Morus alba, 10-30 parts of Poria cocos, 10-30 parts of reed root, 10-30 parts of epimedium, 10-30 parts of raw licorice Weigh 20 parts, 10-30 parts of each Chinese medicine raw material, chop, wash and soak in cold water overnight, then add purified water 15 times the weight of the raw material, water bath to 90 ° C, and keep at 90 ° C, boil for 2 Hours, stirring once every 10 minutes during the cooking process;

[0062] b. Discard the dregs of the traditional Chinese medicine in a, collect the medicinal liquid, and centrifuge at 10,000 rpm for 10 minutes after cooling at room temperature, discard the precipitate, and collect the supernatant;

[0063] c. Precipitate the supernatant in b with alcohol, and obtain the crude Chinese medicine polysaccharide by precipitation;

[...

Embodiment 3

[0067] The preparation of embodiment 3 traditional Chinese medicine flavonoids comprises the following steps:

[0068] a. 10-30 parts of raw land, 20-40 parts of Achyranthes bidentata, 10-20 parts of Morus alba, 10-30 parts of Poria cocos, 10-30 parts of reed root, 10-30 parts of epimedium, 10-30 parts of raw licorice Weigh 20 parts, 10-30 parts of each Chinese medicine raw material, chop, wash and soak in cold water overnight, then add purified water 15 times the weight of the raw material, water bath to 90 ° C, and keep at 90 ° C, boil for 2 Hours, stirring once every 10 minutes during the cooking process;

[0069] b. Discard the dregs of the traditional Chinese medicine in a, collect the medicinal liquid, and centrifuge at 10,000 rpm for 10 minutes after cooling at room temperature, discard the precipitate, and collect the supernatant;

[0070] c. Ethanol precipitation is carried out on the supernatant in b, and the supernatant can obtain the flavonoids of traditional Chin...

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Abstract

The invention discloses a preparation method of a complex live vaccine for porcine reproductive and respiratory syndrome, belonging to the technical field of veterinary biological products. The preparation method comprises the following steps: (1) culturing master cells; (2) performing suspension culture on vaccine preparation cells; (3) culturing and harvesting viruses; (4) performing mixed adsorption on the harvested full-suspension virus solution and chicken anti-porcine reproductive and respiratory syndrome virus egg yolk antibody; (5) adding the adsorbed antigen-antibody compound into an immunoenhancer-containing freeze-drying protecting agent, sub-packaging and then performing freeze vacuum drying to prepare the complex live vaccine for the porcine reproductive and respiratory syndrome. The method is used for preparing the vaccine, and has the characteristics that the production period can be shortened, the personnel allocation is reduced, the pollution probability is low, the floor area is small, the virus titer is high, the difference between the prepared vaccine batches is small, the product has stable and high-efficiency quality, the side effect is small and the quality of the vaccine can be completely improved.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a preparation method of a porcine reproductive and respiratory syndrome compound live vaccine. Background technique [0002] Pig blue ear disease, also known as porcine reproductive and respiratory syndrome (Porcine reproductive and respiratory syndrome, PRRS), is caused by porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome virus, PRRSV). , abortion, stillbirth, weak piglets and mummified fetuses, piglets and fattening pigs with respiratory symptoms. It is difficult to effectively prevent and control PRRS. Firstly, PRRSV targets lung macrophages, which has antibody-dependent enhancement (ADE), causing certain immunosuppression. Secondly, PRRSV is constantly mutating, and there is no effective treatment for PRRS Drugs and immunization are currently the most important measures to control the disease...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/42A61K39/12A61P37/04
Inventor 李阳闫艳丽刘雪王二先徐晓艳盛璐丝
Owner 浙江美保龙生物技术有限公司
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