Preparation method of canine distemper and canine parvovirus bivalent inactivated vaccine

Abstract

The invention belongs to the technical field of preparation of canine distemper and canine parvovirus bivalent inactivated vaccines, and particularly relates to a preparation method of a canine distemper and canine parvovirus bivalent inactivated vaccine, which is characterized by comprising the following steps of: selection and culture of virus strains: selecting a canine distemper virus strain and a canine parvovirus strain which are verified and have stability, and culturing the canine distemper virus strain and the canine parvovirus strain in a cell culture medium, so as to obtain the canine distemper and canine parvovirus bivalent inactivated vaccine. The two virus strains are inoculated respectively, so that the viruses are replicated and proliferated in cells; the bivalent inactivated vaccine for the canine distemper and the canine parvovirus prepared by adopting the preparation method is excellent and stable in efficacy, after the canine is inoculated, no untoward effect is caused, the immune effect is good, the bivalent inactivated vaccine for the canine distemper and the canine parvovirus integrates a canine distemper vaccine and a canine parvovirus vaccine, the canine only needs to be inoculated for one time, and the vaccine is safe and reliable. The canine distemper virus and the canine parvovirus can be immunized at the same time, the preparation method is simple, and the vaccine effect stability is high.

Description

Technical Field

[0001] The present invention belongs to the technical field of the preparation of canine distemper and canine parvovirus bivalent inactivated vaccine, and particularly relates to a preparation method of canine distemper and canine parvovirus bivalent inactivated vaccine. Background Art

[0002] Dogs are widely used as experimental dogs, military dogs, police dogs, pet dogs, etc., but the probability of dying from canine distemper virus (CDV) and canine parvovirus (CPV) is very high, especially when they are young, and these two viruses are infectious among dogs. Once infected, it often costs a lot but with little effect. At present, the quality of canine distemper and canine parvovirus inactivated vaccines on the market is unstable, and there are often defects such as immune failure, short immune time, and many adverse reactions. Summary of the Invention

[0003] To solve the problems raised in the above background art, the present invention provides a preparation method of canine distemper and canine parvovirus bivalent inactivated vaccine, which has the characteristic of stable quality of canine distemper and canine parvovirus inactivated vaccine.

[0004] To achieve the above object, the present invention provides the following technical solution: A preparation method of canine distemper and canine parvovirus bivalent inactivated vaccine, and its preparation method comprises the following steps:

[0005] Selection and culture of virus strains: Select a canine distemper virus strain and a canine parvovirus strain that have been verified and have stability. In a cell culture medium, inoculate these two virus strains respectively, and let the virus replicate and proliferate in the cells;

[0006] Harvest and purification of virus liquid: When the cytopathic effect reaches a predetermined level (such as when the cytopathic effect of cells reaches 80% or more than 90%), harvest the culture solution containing the virus, and remove cell debris and impurities through physical methods such as centrifugation and filtration to purify the virus liquid;

[0007] Virus inactivation: Use a suitable inactivating agent to inactivate the virus liquid. This step is to eliminate the activity of the virus while maintaining its antigenicity. During the inactivation process, the temperature and time need to be strictly controlled to ensure that the virus is completely inactivated without affecting its antigen structure;

[0008] Vaccine formulation: Mix the inactivated canine distemper virus liquid and canine parvovirus liquid in a certain proportion and add an appropriate amount of adjuvant to improve the immune effect and stability of the vaccine;

[0009] Vaccine quality detection: Conduct a series of quality detections on the prepared vaccine, including sterility inspection, safety test, potency test, etc., to ensure that the vaccine does not contain any harmful microorganisms and has good immunogenicity and safety;

[0010] Vaccine Sub - packaging and Preservation: Sub - package the qualified vaccines into appropriate containers, such as syringes or vaccine vials, and store the vaccines under specified conditions to ensure their stability and shelf life.

[0011] Preferably, in the selection and cultivation of virus strains, the cell culture medium can be Vero cells or F81 cell culture medium.

[0012] Preferably, in cell inactivation, the inactivating agent can be formaldehyde solution or β - propiolactone (BPL).

[0013] Preferably, in the selection and cultivation of virus strains, the virus cultivation time is 4 - 5 days, and the temperature is controlled at 35 - 38°C.

[0014] Preferably, in virus inactivation, the inactivation time is 1 - 2 days, and the virus inactivation temperature is controlled at 34 - 37°C.

[0015] Preferably, in vaccine formulation, the adjuvant can be aluminum hydroxide, oil - in - water emulsion adjuvant or liposome adjuvant.

[0016] Preferably, in vaccine sub - packaging and preservation, the syringes or vaccine vials can be stored at 2 - 8°C, and the storage environment should be light - proof and dry, and avoid storing in an environment with peculiar smell and microbial contamination.

[0017] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0018] In the present invention, the canine distemper and canine parvovirus bivalent inactivated vaccine prepared by the preparation method of the present invention has excellent and stable efficacy. After dogs are vaccinated, there are no adverse reactions, and the immune effect is good. The canine distemper and canine parvovirus bivalent inactivated vaccine combines the canine distemper vaccine and the canine parvovirus vaccine into one. Dogs only need to be vaccinated once to be immunized against both canine distemper virus and canine parvovirus at the same time. Moreover, the preparation method in the present invention is simple, and the stability of the vaccine effect is high. Detailed Embodiments

[0019] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0020] Example 1

[0021] The present invention provides the following technical solutions: A preparation method of a canine distemper and canine parvovirus bivalent inactivated vaccine, and its preparation method comprises the following steps:

[0022] Selection and Cultivation of Virus Strains: Select verified and stable canine distemper virus strains and canine parvovirus strains, and inoculate these two virus strains separately in cell culture medium to allow the viruses to replicate and proliferate in the cells;

[0023] Harvest and Purification of Virus Liquid: When the cytopathic effect reaches a predetermined level (such as more than 80% or 90% of the cells showing cytopathic effect), harvest the culture medium containing the virus, and remove cell debris and impurities through physical methods such as centrifugation and filtration to purify the virus liquid;

[0024] Virus Inactivation: Use a suitable inactivating agent to inactivate the virus liquid. This step is to eliminate the activity of the virus while maintaining its antigenicity. During the inactivation process, temperature and time need to be strictly controlled to ensure that the virus is completely inactivated without affecting its antigenic structure;

[0025] Vaccine Preparation: Mix the inactivated canine distemper virus liquid and canine parvovirus liquid in a certain proportion and add an appropriate amount of adjuvant to improve the immune effect and stability of the vaccine;

[0026] Vaccine Quality Detection: Conduct a series of quality detections on the prepared vaccine, including sterility inspection, safety test, and potency test, etc., to ensure that the vaccine does not contain any harmful microorganisms and has good immunogenicity and safety;

[0027] Vaccine Subpackaging and Storage: Subpackage the qualified vaccine into appropriate containers, such as syringes or vaccine bottles, and store the vaccine under specified conditions to ensure its stability and shelf life.

[0028] Specifically, in the selection and cultivation of virus strains, the cell culture medium can be Vero cells or F81 cell culture medium.

[0029] Specifically, in cell inactivation, the inactivating agent can be formaldehyde solution or β-propiolactone (BPL).

[0030] Specifically, in the selection and cultivation of virus strains, the virus cultivation time is 4 - 5 days, and the temperature is controlled at 35 - 38°C.

[0031] Specifically, in virus inactivation, the inactivation time is 1 - 2 days, and the virus inactivation temperature is controlled at 34 - 37°C.

[0032] Specifically, in vaccine preparation, the adjuvant can be aluminum hydroxide, oil emulsion adjuvant, or liposome adjuvant.

[0033] Specifically, in vaccine subpackaging and storage, the syringe or vaccine bottle can be stored at 2 - 8°C, and the storage environment should be light-proof and dry, and avoid storing in an environment with peculiar smell and microbial contamination.

[0034] Finally, it should be noted that the above are only preferred embodiments of the present invention and are not used to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or perform equivalent replacements for some of the technical features. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A method for preparing a canine distemper and canine parvovirus combined inactivated vaccine, characterized in that: The preparation method thereof comprises the following steps: Selection and cultivation of virus strains: Select verified and stable canine distemper virus strains and canine parvovirus strains, inoculate these two virus strains separately in cell culture medium, and allow the viruses to replicate and proliferate in cells; Harvest and purification of virus fluid: When the cell lesion reaches a predetermined level (such as cell lesion reaches 80% or 90% or more), harvest the culture fluid containing the virus, remove cell debris and impurities through physical methods such as centrifugation and filtration, and purify the virus fluid; Virus inactivation: Use a suitable inactivator to inactivate the virus solution. This step is to eliminate the activity of the virus while maintaining its antigenicity. The temperature and time must be strictly controlled during the inactivation process to ensure that the virus is completely inactivated without affecting its antigenic structure. Vaccine preparation: Mix the inactivated canine distemper virus liquid and canine parvovirus liquid in a certain proportion and add an appropriate amount of adjuvant to improve the pain-free effect and stability of the vaccine; Vaccine quality testing: A series of quality tests are conducted on the prepared vaccines, including sterility tests, safety tests, and efficacy tests, to ensure that the vaccines do not contain any harmful microorganisms and have good immunogenicity and safety; Vaccine packaging and storage: Vaccines that pass the test should be packaged into appropriate containers, such as syringes or vaccine bottles, and stored under specified conditions to ensure their stability and shelf life.

2. The method for preparing the canine distemper and canine parvovirus combined inactivated vaccine according to claim 1, characterized in that: In the selection and cultivation of virus strains, the cell culture medium can be selected from Vero cell or F81 cell culture medium.

3. The method for preparing the canine distemper and canine parvovirus combined inactivated vaccine according to claim 1, characterized in that: In cell inactivation, the extinguishing agent can be formaldehyde solution or beta-propiolactone (BPL).

4. The method for preparing the canine distemper and canine parvovirus combined inactivated vaccine according to claim 1, characterized in that: In the selection and cultivation of virus strains, the virus cultivation time is 4-5 days and the temperature is controlled at 35-38°C.

5. The method for preparing the canine distemper and canine parvovirus combined inactivated vaccine according to claim 1, characterized in that: In virus inactivation, the inactivation time is 1-2 days, and the virus inactivation temperature is controlled at 34-37°C.

6. The method for preparing the canine distemper and canine parvovirus combined inactivated vaccine according to claim 1, characterized in that: In vaccine formulation, adjuvants can be aluminum hydroxide, oil emulsion adjuvants or liposome adjuvants.

7. The method for preparing the canine distemper and canine parvovirus combined inactivated vaccine according to claim 1, characterized in that: During vaccine packaging and storage, syringes or vaccine bottles can be stored at 2-8°C in a light-proof, dry environment, and avoid storage in an environment with odor or microbial contamination.