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Preparation method of PCV2, PRRSV and mycoplasma hyopneumoniae triple inactivated vaccine

A technology of Mycoplasma hyopneumoniae and inactivated vaccine, applied in the field of veterinary biological products, can solve problems such as shortage, and achieve the effects of ensuring uniformity, improving immune effect and production performance, and improving stability

Inactive Publication Date: 2018-03-06
TIANJIN RINGPU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims at the technical defect of prior art, provides a kind of PCV2, PRRSV, mycoplasma hyopneumoniae triple inactivated vaccine preparation method, to solve the lack of a kind of vaccine that can simultaneously immunize porcine circovirus type 2, porcine reproductive Technical Problems of Vaccines Against Respiratory Syndrome Virus and Porcine Mycoplasma Pneumonia

Method used

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  • Preparation method of PCV2, PRRSV and mycoplasma hyopneumoniae triple inactivated vaccine
  • Preparation method of PCV2, PRRSV and mycoplasma hyopneumoniae triple inactivated vaccine
  • Preparation method of PCV2, PRRSV and mycoplasma hyopneumoniae triple inactivated vaccine

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Experimental program
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Effect test

Embodiment 1

[0034] A preparation method of PCV2, PRRSV, mycoplasma hyopneumoniae triple inactivated vaccine, comprising the following steps:

[0035] 1. Production seed preparation

[0036] 1.1 Porcine circovirus production seed preparation

[0037] 1.1.1 Cell Propagation

[0038] Resuscitate PK15 cells, add cell nutrient solution, culture at 37°C for 24-48 hours, and pass passage after forming a good cell monolayer. When passaging, discard the nutrient solution, add EDTA-trypsin digestion solution, act for several minutes, digest and disperse evenly to make cell suspension. Then it was divided into cell culture flasks, incubated at 37°C for 24 hours, and tested for sterility, mycoplasma and exogenous virus according to the appendix of the current "Chinese Veterinary Pharmacopoeia", which should meet the requirements.

[0039] 1.1.2 Breeding of poisonous seeds

[0040] Inoculate PK-15 cells into cell culture medium, adjust the concentration to 200,000-300,000 / mL, and incubate with PCV...

Embodiment 2

[0080] The safety research of the vaccine prepared in embodiment 2 embodiment 1

[0081] Get the vaccine in Example 1, mark the head portion (2mL / head portion) by the bottle label, intramuscularly inject 5 portions after weaning 1.5-2 months old pigs (negative swine fever neutralizing antibody, pig PCV2ELISA antibody negative, pig PRRSVELISA Antibody negative, Mycoplasma swine ELISA antibody negative) 5 heads, 5 heads in the control group were not immunized, observed daily for 21 days, there should be no local or systemic adverse reactions caused by the injection of the vaccine. The results are shown in Table 1.

[0082] Table 1 Safety study results

[0083]

[0084] The safety test results showed that the triple inactivated vaccine of porcine circovirus type 2, porcine reproductive and respiratory syndrome virus and mycoplasma hyopneumoniae was safe for weaned piglets.

Embodiment 3

[0085] Example 3 Vaccine Vaccine Efficacy Research Prepared in Example 1

[0086] 1.1 Porcine circovirus type 2 vaccine immunization and challenge

[0087] Fifteen healthy susceptible piglets with negative PCV2 ELISA antibody, PCV2 antigen negative and PRRSV ELISA antibody negative (commercially registered ELISA antibody kit detection) aged 14-21 days were randomly divided into 3 groups, 5 pigs in each group. The first group is the triple inactivated vaccine immunization group of porcine circovirus type 2, porcine reproductive and respiratory syndrome virus, and mycoplasma hyopneumoniae, intramuscularly injecting vaccine 2mL / head; the second group is the PCV2 vaccine immunization group, and the third group is non-inactivated Blank control was inoculated. The virus was challenged 28 days after vaccination, and each pig was weighed before the challenge. Group 1 was inoculated with PCV2 (10 7.0 TCID 50 4.0 mL / ml), wherein each pig was intranasally dripped with 4.0 mL, and the...

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Abstract

The invention provides a preparation method of a PCV2, PRRSV and mycoplasma hyopneumoniae triple inactivated vaccine. The technical scheme is that a brand-new preparation method is designed from culture, inactivation, complex and other levels based on growth characteristics of three microorganisms and physicochemical characteristics of an antigen substance. In the method, the PCV2 and the PRRSV are prepared by adopting a bioreactor mode; in the preparation process of an antigen, a pH value of fermented liquid is strictly controlled, so that the immunogenicity of a virus is effectively ensured;mycoplasam suis produces a strain in a bacteria fermenting tank; and in the breeding process of the strain, the homogenization of an intra-batch bacterial solution is ensured by setting dissolved oxygen, the pH value, rotating speed and the like, the defects of complex operation, high working intensity, easiness in pollution and the like in the rolling bottle culture process are eliminated, and the quality stability of the mycoplasam suis inactivated vaccine is effectively improved. Experiments show that the protection rates of the prepared triple inactivated vaccine for the PCV2, the PRRSV and the mycoplasma hyopneumoniae all exceed 80 percent, and preventing and treating demands on three pathogens are met.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, and further relates to the preparation technology of multiple vaccines, in particular to a preparation method of triple inactivated vaccine of PCV2, PRRSV and Mycoplasma hyopneumoniae. Background technique [0002] Porcine circovirus type 2 (PCV2) is believed to be the cause of multisystemic wasting and wasting syndrome (PMWS) in weaned pigs characterized by severe wasting, lymphadenopathy, and respiratory symptoms. The main characteristics of porcine circovirus infection are physical decline, weight loss, anemia, jaundice, poor growth and development, diarrhea, dyspnea, general fingering disorder of sows, extensive lesions of internal organs and skin, especially kidney, spleen and systemic lymph nodes Hemorrhage, necrosis and other symptoms bring huge economic losses to the pig industry. Mixed infections of PCV2 and other pathogens are often reported, such as porcine repr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61K39/02A61K39/295A61P31/20A61P31/14A61P31/04
CPCA61K39/0241A61K39/12A61K2039/521A61K2039/5252A61K2039/552A61K2039/70C12N2750/10034C12N2770/10034A61K2300/00
Inventor 厚华艳吕茂杰付旭彬郁宏伟梁武李守军
Owner TIANJIN RINGPU BIO TECH
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