Inactivation inspection method suitable for BVDV, IBRV, PIV3 and BRSV inactivated vaccines

A technology of inactivation testing and inactivation vaccines, which is applied in the fields of biomedicine and veterinary medicine to achieve the effects of high detection rate, high accuracy and improved safety

Inactive Publication Date: 2018-06-12
华威特(江苏)生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, the inactivation test method of porcine circovirus inactivated vaccine is not suitable for the inactivation test of bovine viral diarrhea virus and bovine infectious rhinotracheitis virus

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The inactivation test method of embodiment 1 bovine viral diarrhea virus inactivated vaccine

[0020] Cells and cell culture medium: MDBK cells, cell culture medium is DMEM nutrient solution containing 6% newborn bovine serum; cell maintenance medium is DMEM nutrient solution containing 3% horse serum.

[0021] 1) Inoculate the digested MDBK cells into a 3000ml spinner bottle and culture for 48 hours until 80%-90% good cell monolayers grow, discard the cell culture medium, and take the bovine viral diarrhea virus liquid cultured by the inactivated MDBK cells 25ml was inoculated in the cell spinner bottle, placed at 37°C, containing 5% CO 2 In the incubator, make the inactivated virus liquid adsorb on suitable cells for 40 minutes, and shake twice during the period;

[0022] 2) After adsorption, add 175ml of DMEM cell maintenance solution containing 3% horse serum to the cell flask, and place the cell flask at 37°C in CO 2 After continuing to culture in the incubator f...

Embodiment 2

[0025] Example 2 Inactivation test method of bovine infectious rhinotracheitis cytotoxic inactivated vaccine

[0026] Cells and cell culture medium: MDBK cells or BT cells, the cell culture medium is MEM nutrient solution containing 6% newborn bovine serum; the cell maintenance medium is MEM nutrient solution containing 3% newborn bovine serum.

[0027] 1) Inoculate digested MDBK cells into a 3000ml spinner bottle and culture for 48 hours until 80%-90% of good cell monolayers grow, discard the cell culture medium, and take inactivated MDBK cells to culture bovine infectious rhinotracheitis Inoculate 25ml of the virus solution into a cell spinner bottle and store at 37°C with 5% CO 2 In the incubator, make the inactivated virus liquid adsorb on suitable cells for 40 minutes, and shake twice during the period;

[0028] 2) After adsorption, add 175ml of DMEM cell maintenance solution containing 3% horse serum to the cell flask, and place the cell flask at 37°C in CO 2 After con...

Embodiment 3

[0031] Example 3 The inactivation test method of bovine parainfluenza type 3 cytotoxic inactivated vaccine

[0032] Cells and cell culture medium: MDBK cells or BT cells, the cell culture medium is MEM nutrient solution containing 6% newborn bovine serum; the cell maintenance medium is MEM nutrient solution containing 3% newborn bovine serum.

[0033] 1) Inoculate digested MDBK cells into a 3000ml spinner bottle and culture for 48 hours until 80%-90% of good cell monolayers grow, discard the cell culture medium, and take inactivated MDBK cells to culture bovine infectious rhinotracheitis Inoculate 25ml of the virus solution into a cell spinner bottle and store at 37°C with 5% CO 2 In the incubator, make the inactivated virus liquid adsorb on suitable cells for 40 minutes, and shake twice during the period;

[0034] 2) After adsorption, add 175ml of DMEM cell maintenance solution containing 3% horse serum to the cell flask, and place the cell flask at 37°C in CO 2 After conti...

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PUM

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Abstract

The invention discloses an inactivation inspection method suitable for BVDV, IBRV, PIV3 and BRSV inactivated vaccines, and relates to the field of biological medicines, in particular to an inactivation inspection method for BVDV, IBRV, PIV3 and BRSV inactivated vaccines. The inactivation inspection method comprises the following steps: (1) adsorbing an inactivated virus liquor on an appropriate cell for 40-60 min, and shaking for 2-3 times in the period; (2) after adsorption, adding a certain volume of a cell maintenance fluid in a cell spinner bottle, continuously performing culture, observing cytopathy, as for the virus fluid free from cytopathy, harvesting a cell culture fluid, and performing freeze thawing for 2-3 times; and (3) taking 10-15 ml of the frozen and thawed cell culture fluid, inoculating the taken cell culture fluid on the appropriate cell for adsorption, and continuously performing culture. Observation and fluorescence detection are performed, and if no specific fluorescence appears, complete inactivation is manifested. According to the technical scheme disclosed by the prevention, the anti-gen concentration and the virus infection time are guaranteed. The virus detection rate of the inactivation inspection method is high, and the bio-safety of the inactivated vaccines can be effectively ensured.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to an inactivated vaccine suitable for bovine viral diarrhea (BVDV), bovine infectious rhinotracheitis (IBRV), bovine parainfluenza (PIV3) and bovine respiratory syncytial virus disease (BRSV) The inactivation test method belongs to the field of veterinary medicine and biology. Background technique [0002] Vaccine immunization is an important means of preventing and controlling epidemic diseases. According to whether the pathogenic microorganisms in the vaccine survive, vaccines are divided into inactivated vaccines and live vaccines. Inactivated vaccines are pathogenic microorganisms that use chemical or physical methods to eliminate the activity, fecundity or pathogenicity of the microorganisms used to make vaccines, but still retain the immunogenicity of the corresponding antigens. [0003] Inactivation test is one of the important processes in the preparation of inactivated vaccines...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569
CPCG01N33/56983G01N33/577
Inventor 王炜师新川李真光孟庆森黄慧文刘芳芳季烨武华
Owner 华威特(江苏)生物制药有限公司
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