CN105833262A - Porcine circovirus II type inactivated vaccine composition and preparation method - Google Patents
Porcine circovirus II type inactivated vaccine composition and preparation method Download PDFInfo
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Abstract
The invention discloses a porcine circovirus II type inactivated vaccine composition and a preparation method to solve the problem in the prior art that a porcine circovirus II type vaccine is high in cost and low in effect. The porcine circovirus II type inactivated vaccine composition consists of the following components: a porcine circovirus II type inactivated virus bulk, a Chinese herbal medicine adjuvant, and a chemical adjuvant, wherein the Chinese herbal medicine adjuvant consists of ginseng, eucommia bark and angelica polysaccharide, the chemical adjuvant consists of white oil, tween-20, span-80, and aluminum stearate, and the mass ratio of the porcine circovirus II type strain inactivated virus bulk to the Chinese herbal medicine adjuvant is 1:2-2:1. The invention also discloses a method for preparing a vaccine. In the method, the Chinese herbal medicine adjuvant playing an immune role and the porcine circovirus II type inactivated virus bulk are mixed uniformly to ensure that the vaccine is higher in effect and low in cost; and after an antigen substance is mixed with the adjuvants to be injected into an organism, the physical property of the antigen is changed, so that the antigen substance can be released slowly, and the acting time of the antigen is prolonged.
Description
Technical field
The present invention relates to pig vaccines arts, particularly relate to a kind of porcine circovirus type II inactivated vaccine compositions
And preparation method.
Background technology
Porcine circovirus 2 type (PCV2) is the master causing pig postweaning multisystem exhaustion syndrome (PMWS)
Want cause of disease.Since within 1991, breaking out in Canada swinery, PMWS causes to whole world pig industry
Huge economic loss.PCV2 is also scorching with Hypertrophic necrotizing pneumonia, sow breeding difficulty, Corii Sus domestica and nephropathy
Syndrome is relevant with porcine respiratory disease complex.According to estimates, PCV2 diseases related (PCVD), mainly
Being PMWS, the loss caused to Europe in 2014 is up to 600,000,000 Euros, and PCVD raises pigs to China equally
Industry causes serious economic loss.
At present, traditional PCV2 vaccine is generally by inactivateing or being killed or modified pathogenic microorganism alive
Thick prepared product composition, adds the adjuvant of smaller dose in vaccine, stimulates similar immunne response, thus
Reduce the manufacturing cost of vaccine.Therefore, can dramatically increase, when by vaccine with adjuvant combination, the effect vaccinated
Power.
Summary of the invention
The present invention proposes a kind of porcine circovirus type II inactivated vaccine compositions and preparation method, solves existing
The problem that in technology, porcine circovirus type 2 vaccines cost is high, effect is low.
The technical scheme is that and be achieved in that: a kind of porcine circovirus type II inactivated vaccine compositions,
It is characterized in that, described compositions is composed of the following components: pig gyrate virus II type inactivation of viruses liquid, medium-height grass
Medicine adjuvant, chemical classes adjuvant, wherein said Chinese Herbal Medicines Adjuvant is by astragalus polysaccharides, Radix Ginseng, Folium Eucommiae, Radix Angelicae Sinensis
Polysaccharide forms, and described chemical classes adjuvant is white oil, polysorbas20, sorbester p17, stearic acid aluminum, described pig annulus
Virus type II strain inactivation of viruses liquid is 1:2～2:1 with the mass ratio of described Chinese Herbal Medicines Adjuvant.
As preferred technical scheme, described compositions comprises the raw material of following weight portion:
20～25 parts of astragalus polysaccharidess, 10～15 parts of Radix Ginsengs, 3～6 parts of Folium Eucommiaes, 20～30 parts of Radix Angelicae Sinensis polysaccharides, 30～50
Part pig gyrate virus II type inactivation of viruses liquid, 18～24 parts of white oils, 12～15 parts of polysorbas20s, 6～10 parts of span
80,4～7 parts of stearic acid aluminum.
As preferred technical scheme, the malicious valency of described pig gyrate virus II type inactivation of viruses liquid is
105.5TCID50/mL。
The effect on vaccine that selects of immunological adjuvant affects very greatly, and it both can change immunoreation type, makes body
Liquid immunity is changed into cellular immunization, changes kind and the subclass of immunoglobulin, also can change immunoreation shape
State, eliminates immunologic tolerance and produces immunoreation.Therefore, suitable immunological adjuvant is selected to seem the heaviest
Want.
The Chinese Herbal Medicines Adjuvant of the present invention selects astragalus polysaccharides, Radix Ginseng, Folium Eucommiae, Radix Angelicae Sinensis polysaccharide, astragalus polysaccharides
The immunity of body is mainly improved by the proliferation and differentiation of stimulator antigen presenting cell and the release cells factor,
Active component in Radix Ginseng is saponin, the proliferation and differentiation of saponin energy stimulator antigen presenting cell and release cells because of
Son improves the immunity of body.Folium Eucommiae is benefit vital essence class Chinese herbal medicine, can eliminate immunity of organism toleration and
Producing immunoreation, Radix Angelicae Sinensis polysaccharide is made up of glucose, arabinose and a small amount of alditol, and Radix Angelicae Sinensis polysaccharide is not
Directly act on lymphocyte, but first act on nonspecific immunity cell such as DC and make it obtain
Activation, function strengthen, and remake for various immune effector cells.
Stearic acid aluminum as adjuvant, its mechanism of action: after sticking antigen, change antigen physical behavior,
Form storehouse effect, extend antigen presenting cell (APC) to the picked-up of antigen, time of processing, processing, make
Immunocyte keeps long-time high-caliber response to this antigen.
Polysorbas20 is aqueous phase, and sorbester p17 is emulsifying agent, and white oil is as oil phase, and its mechanism of action is antigen bag
By in the micro structure that oil phase is formed, it is allowed to form depots and slowly discharge, stimulates bone-marrow-derived lymphocyte to produce
Antibody.
The present invention proposes again the preparation method of a kind of porcine circovirus type II inactivated vaccine compositions, including following
Step:
1) active component of Chinese Herbal Medicines Adjuvant is extracted, afterwards with physiological saline solution, make after filtration and treat
Use liquid；
2) prepare pig gyrate virus II type inactivation of viruses liquid, described pig gyrate virus II type inactivation of viruses liquid is added
To described stand-by liquid, stir；
3) in white oil, add sorbester p17 and stearic acid aluminum makes oil mixture, using the tween as aqueous phase
20 mix with described oil mixture, obtain mixed liquor；
4) by step 2) in product join in described mixed liquor, centrifugal emulsifying 3min, get product.
As preferred technical scheme, step 2) in prepare pig gyrate virus II type inactivation of viruses liquid during
Including designing three to primer for expanding the complete genome sequence of pig gyrate virus II type strain, described three pairs of primers
Respectively:
The gene order of PCV2-F1 is 5'-TGCAGAAGCGTGATTGGAAGAC-3',
The gene order of PCV2-R1 is 5'-CCGCACCTTCGGATATACTATC-3'；
The gene order of PCV2-F2 is 5'-AGGGCCAGAATTCAACCTTT-3',
The gene order of PCV2-R2 is 5'-TCCCACAGAGTCTCAGTAGA-3'；
The gene order of PCV2-F3 is 5'-AGGGCCAGAATTCAACCTTAACCTTC-3',
The gene order of PCV2-R3 is 5'-GTTGAATTCTGGCCCTGCTCCCCGAT-3'.
The present invention proposes the method preparing vaccine, will play Chinese Herbal Medicines Adjuvant and the pig circular ring virus 2 of immunization
Poison II type inactivation of viruses liquid mix homogeneously so that the effect of vaccine is higher, low cost, in order to allow stearic acid aluminum
Preferably it is dissolved in white oil, adds the sorbester p17 as emulsifying agent, the most again white oil is mixed with polysorbas20,
Form water-in-oil type adjuvant, mix with antigen, traditional Chinese medicine adjuvant, after antigenic substance mixing adjuvant injects body,
Change the physical property of antigen, antigenic substance can be made to discharge slowly, extend the action time of antigen.
Accompanying drawing explanation
In order to be illustrated more clearly that the embodiment of the present invention or technical scheme of the prior art, below will be to enforcement
In example or description of the prior art, the required accompanying drawing used is briefly described, it should be apparent that, describe below
In accompanying drawing be only some embodiments of the present invention, for those of ordinary skill in the art, do not paying
On the premise of going out creative work, it is also possible to obtain other accompanying drawing according to these accompanying drawings.
Fig. 1 is PCR primer electrophoresis result；
Fig. 2 is PCV2 full genome H amplification figure；
Fig. 3 is PCV2 full genome H amplification figure.
Detailed description of the invention
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clearly
Chu, be fully described by, it is clear that described embodiment be only a part of embodiment of the present invention rather than
Whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art are not making creation
The every other embodiment obtained under property work premise, broadly falls into the scope of protection of the invention.
A kind of porcine circovirus type II inactivated vaccine compositions, it is characterised in that described compositions is by following group
Be grouped into: pig gyrate virus II type inactivation of viruses liquid, Chinese Herbal Medicines Adjuvant, chemical classes adjuvant, wherein said in
Medical herbs adjuvant is white oil by astragalus polysaccharides, Radix Ginseng, Folium Eucommiae, Radix Angelicae Sinensis polysaccharide, described chemical classes adjuvant, is told
Temperature 20, sorbester p17, stearic acid aluminum, described pig gyrate virus II type strain inactivation of viruses liquid and described Chinese herbal medicine
The mass ratio of adjuvant is 1:2～2:1.
The preferred described pig gyrate virus II type strain inactivation of viruses liquid of the present invention and the quality of described Chinese Herbal Medicines Adjuvant
Ratio is 1:1.
As preferred technical scheme, described compositions comprises the raw material of following weight portion:
20～25 parts of astragalus polysaccharidess, 10～15 parts of Radix Ginsengs, 3～6 parts of Folium Eucommiaes, 20～30 parts of Radix Angelicae Sinensis polysaccharides, 30～50
Part pig gyrate virus II type inactivation of viruses liquid, 18～24 parts of white oils, 12～15 parts of polysorbas20s, 6～10 parts of span
80,4～7 parts of stearic acid aluminum.
As preferred technical scheme, the malicious valency of described pig gyrate virus II type inactivation of viruses liquid is
105.5TCID50/m L。
The present invention proposes again the preparation method of a kind of porcine circovirus type II inactivated vaccine compositions, including following
Step:
1) active component of Chinese Herbal Medicines Adjuvant is extracted, afterwards with physiological saline solution, make after filtration and treat
Use liquid；
2) prepare pig gyrate virus II type inactivation of viruses liquid, described pig gyrate virus II type inactivation of viruses liquid is added
To described stand-by liquid, stir；
3) in white oil, add sorbester p17 and stearic acid aluminum makes oil mixture, using the tween as aqueous phase
20 mix with described oil mixture, obtain mixed liquor；
4) by step 2) in product join in described mixed liquor, centrifugal emulsifying 3min, get product.
The present invention prepare pig gyrate virus II type virus concrete grammar:
1, virus purification is cultivated
Taking doubtful PCV2 infected pigs lungs or lymph node, add DMEM and grind, 0.22 μm membrane filtration removes
Bacterium is inoculated in monolayer PK-15 cell, abandons supernatant after hatching 1h, continues to cultivate, blind passage 20 generation.Repeatedly freeze
Melt cell and collect virus.
2, for design of primers and the synthesis of whole genome amplification
Design three to primer PCV2-F1/R1, PCV2F2/R2 and PCV2F3/R3 tri-to primer (being shown in Table 1)
For expanding PCV2 full-length genome.
Table 1 primer and sequence
3, viral DNA extracts
Extracting test kit operation instructions according to viral DNA to operate, step is as follows:
Take 300 μ L cell suspension and add the BufferPLY of 2 times of volumes.Mix homogeneously, room temperature places 15min.
The dehydrated alcohol of 0.5 times of volume is added, reverse five mixings in mixed liquor.Upper step mixed liquor is proceeded to ezBind
Columns, 12000r/min are centrifuged 1min.Adding 650 μ L Wash Buffer, 12000r/min is centrifuged 1min,
It is repeated once.Discarding washing liquid, idle running ezBind, 12000r/min are centrifuged 2min, are shifted by ezHind post
To clean eppendorf pipe, adding 15 μ L ddH2O, it is centrifugal that room temperature places 2min, 12000r/min
2min, gathers in the crops DNA.
4, purpose nucleotide cloning and sequencing
The step for include the recovery of PCR primer, connection, the preparation of DH5a competent escherichia coli cell,
Connect that product carries out converting, the extraction of plasmid DNA, the double digestion of recombiant plasmid are identified, positive colony sequence
Measure.
5, result
5.1 PCV2-SD nucleic acid fragment amplifications
The STb gene extracted in tissue, as template, applies two specific primer carries out PCR amplification respectively,
After agarose gel electrophoresis, obtain 1057bp (A) and the nucleic acid fragment of 1110bp (B) respectively, with expection
In the same size, result is shown in Fig. 1.
With fragment A, B mixture as template, with PCV2-F3/R3 as primer, carry out PCR reaction,
To PCV2 full-length genome H, clip size 1767bp, such as Fig. 2.
The clone of 5.2 PCR primer and the qualification of recombiant plasmid
After the PCR primer of amplification is connected with pMD 18-T carrier, connects product and convert escherichia coli DH5a
Competent cell, the plasmid of extraction Sal I and BamH I enzyme action, electrophoresis result such as Fig. 3.Recombiant plasmid
After enzyme action, obtain the nucleic acid fragment of 1767bp and 2692bp, in the same size with expection.
The method preparing pig gyrate virus II type inactivation of viruses liquid:
Being cultivated by PCV2 strain inoculation PK15 cell, viral cultures gather in the crops after 3 freeze thawing, through high speed from
The heart abandons cell impurities, then through the suitable concentrating virus of ultracentrifugation, puts-70 DEG C of Refrigerator stores standby.
Measure PCV2-SD concentrating virus liquid poison valency, be adjusted to 105.5TCID50/m L, add formaldehyde, make
The ultimate density of formaldehyde is 0.1%.37 DEG C of constant-temperature table inactivation 18h.Extract respectively a small amount of inactivation of viruses liquid with
It is inoculated in monolayer PK15 cell after 1:10 dilution, in blind passage 3 generation, to obtain final product.
The present invention illustrates the invention with specific embodiment.
Embodiment one
The preparation method of a kind of porcine circovirus type II inactivated vaccine compositions, comprises the following steps:
1) by 20 parts of astragalus polysaccharidess, 10 parts of Radix Ginsengs, 3 parts of Folium Eucommiaes, 20 Radix Angelicae Sinensis polysaccharide Chinese Herbal Medicines Adjuvants
Active component extracts, and afterwards with physiological saline solution, makes stand-by liquid after filtration；
2) prepare pig gyrate virus II type inactivation of viruses liquid, described pig gyrate virus II type inactivation of viruses liquid is added
To described stand-by liquid, and mix with 1:2 ratio, stir；
3) in 18 parts of white oils, add 6 parts of sorbester p17s and 4 parts of stearic acid aluminum make oil mixture, will make
Mix with described oil mixture for the polysorbas20s of 12 parts of aqueous phase, obtain mixed liquor；
4) by step 2) in product join in described mixed liquor, centrifugal emulsifying 3min, get product.
Embodiment two
The preparation method of a kind of porcine circovirus type II inactivated vaccine compositions, comprises the following steps:
1) by 25 parts of astragalus polysaccharidess, 15 parts of Radix Ginsengs, 6 parts of Folium Eucommiaes, 30 Radix Angelicae Sinensis polysaccharide Chinese Herbal Medicines Adjuvants
Active component extracts, and afterwards with physiological saline solution, makes stand-by liquid after filtration；
2) prepare pig gyrate virus II type inactivation of viruses liquid, described pig gyrate virus II type inactivation of viruses liquid is added
To described stand-by liquid, and mix with 1:2 ratio, stir；
3) in 24 parts of white oils, add 10 parts of sorbester p17s and 7 parts of stearic acid aluminum make oil mixture, will
Mix with described oil mixture as the polysorbas20s of 15 parts of aqueous phase, obtain mixed liquor；
4) by step 2) in product join in described mixed liquor, centrifugal emulsifying 3min, get product.
Embodiment three
The preparation method of a kind of porcine circovirus type II inactivated vaccine compositions, comprises the following steps:
1) by 20 parts of astragalus polysaccharidess, 10 parts of Radix Ginsengs, 3 parts of Folium Eucommiaes, 20 Radix Angelicae Sinensis polysaccharide Chinese Herbal Medicines Adjuvants
Active component extracts, and afterwards with physiological saline solution, makes stand-by liquid after filtration；
2) prepare pig gyrate virus II type inactivation of viruses liquid, described pig gyrate virus II type inactivation of viruses liquid is added
To described stand-by liquid, and mix with 1:1 ratio, stir；
3) in 18 parts of white oils, add 6 parts of sorbester p17s and 4 parts of stearic acid aluminum make oil mixture, will make
Mix with described oil mixture for the polysorbas20s of 12 parts of aqueous phase, obtain mixed liquor；
4) by step 2) in product join in described mixed liquor, centrifugal emulsifying 3min, get product.
Embodiment four
The preparation method of a kind of porcine circovirus type II inactivated vaccine compositions, comprises the following steps:
1) by 25 parts of astragalus polysaccharidess, 15 parts of Radix Ginsengs, 6 parts of Folium Eucommiaes, 30 Radix Angelicae Sinensis polysaccharide Chinese Herbal Medicines Adjuvants
Active component extracts, and afterwards with physiological saline solution, makes stand-by liquid after filtration；
2) prepare pig gyrate virus II type inactivation of viruses liquid, described pig gyrate virus II type inactivation of viruses liquid is added
To described stand-by liquid, and mix with 1:1 ratio, stir；
3) in 24 parts of white oils, add 10 parts of sorbester p17s and 7 parts of stearic acid aluminum make oil mixture, will
Mix with described oil mixture as the polysorbas20s of 15 parts of aqueous phase, obtain mixed liquor；
4) by step 2) in product join in described mixed liquor, centrifugal emulsifying 3min, get product.
Embodiment five
The preparation method of a kind of porcine circovirus type II inactivated vaccine compositions, comprises the following steps:
1) by 20 parts of astragalus polysaccharidess, 10 parts of Radix Ginsengs, 3 parts of Folium Eucommiaes, 20 Radix Angelicae Sinensis polysaccharide Chinese Herbal Medicines Adjuvants
Active component extracts, and afterwards with physiological saline solution, makes stand-by liquid after filtration；
2) prepare pig gyrate virus II type inactivation of viruses liquid, described pig gyrate virus II type inactivation of viruses liquid is added
To described stand-by liquid, and mix with 2:1 ratio, stir；
3) in 18 parts of white oils, add 6 parts of sorbester p17s and 4 parts of stearic acid aluminum make oil mixture, will make
Mix with described oil mixture for the polysorbas20s of 12 parts of aqueous phase, obtain mixed liquor；
4) by step 2) in product join in described mixed liquor, centrifugal emulsifying 3min, get product.
Embodiment six
The preparation method of a kind of porcine circovirus type II inactivated vaccine compositions, comprises the following steps:
1) by 25 parts of astragalus polysaccharidess, 15 parts of Radix Ginsengs, 6 parts of Folium Eucommiaes, 30 Radix Angelicae Sinensis polysaccharide Chinese Herbal Medicines Adjuvants
Active component extracts, and afterwards with physiological saline solution, makes stand-by liquid after filtration；
2) prepare pig gyrate virus II type inactivation of viruses liquid, described pig gyrate virus II type inactivation of viruses liquid is added
To described stand-by liquid, and mix with 2:1 ratio, stir；
3) in 24 parts of white oils, add 10 parts of sorbester p17s and 7 parts of stearic acid aluminum make oil mixture, will
Mix with described oil mixture as the polysorbas20s of 15 parts of aqueous phase, obtain mixed liquor；
4) by step 2) in product join in described mixed liquor, centrifugal emulsifying 3min, get product.
The foregoing is only presently preferred embodiments of the present invention, not in order to limit the present invention, all at this
Within bright spirit and principle, any modification, equivalent substitution and improvement etc. made, should be included in this
Within bright protection domain.
Claims (5)
1. a porcine circovirus type II inactivated vaccine compositions, it is characterised in that described compositions is by following
Component forms: pig gyrate virus II type inactivation of viruses liquid, Chinese Herbal Medicines Adjuvant, chemical classes adjuvant, wherein said
Chinese Herbal Medicines Adjuvant is made up of astragalus polysaccharides, Radix Ginseng, Folium Eucommiae, Radix Angelicae Sinensis polysaccharide, and described chemical classes adjuvant is white
Oil, polysorbas20, sorbester p17, stearic acid aluminum, described pig gyrate virus II type strain inactivation of viruses liquid is with described
The mass ratio of Chinese Herbal Medicines Adjuvant is 1:2～2:1.
2. a kind of porcine circovirus type II inactivated vaccine compositions as claimed in claim 1, it is characterised in that
Described compositions comprises the raw material of following weight portion:
20～25 parts of astragalus polysaccharidess, 10～15 parts of Radix Ginsengs, 3～6 parts of Folium Eucommiaes, 20～30 parts of Radix Angelicae Sinensis polysaccharides, 30～50
Part pig gyrate virus II type inactivation of viruses liquid, 18～24 parts of white oils, 12～15 parts of polysorbas20s, 6～10 parts of span
80,4～7 parts of stearic acid aluminum.
3. a kind of porcine circovirus type II inactivated vaccine compositions as claimed in claim 2, it is characterised in that:
The malicious valency of described pig gyrate virus II type inactivation of viruses liquid is 105.5TCID50/m L。
4. a kind of porcine circovirus type II inactivated vaccine group of preparation
The method of compound, it is characterised in that comprise the following steps:
1) active component of Chinese Herbal Medicines Adjuvant is extracted, afterwards with physiological saline solution, make after filtration and treat
Use liquid；
2) prepare pig gyrate virus II type inactivation of viruses liquid, described pig gyrate virus II type inactivation of viruses liquid is added
To described stand-by liquid, stir；
3) in white oil, add sorbester p17 and stearic acid aluminum makes oil mixture, using the tween as aqueous phase
20 mix with described oil mixture, obtain mixed liquor；
4) by step 2) in product join in described mixed liquor, centrifugal emulsifying 3min, get product.
The preparation method of a kind of porcine circovirus type II inactivated vaccine compositions the most as claimed in claim 4,
It is characterized in that, step 2) in prepare pig gyrate virus II type inactivation of viruses liquid during to include designing three right
Primer for expanding the complete genome sequence of pig gyrate virus II type strain, described three pairs of primers respectively:
The gene order of PCV2-F1 is 5'-TGCAGAAGCGTGATTGGAAGAC-3',
The gene order of PCV2-R1 is 5'-CCGCACCTTCGGATATACTATC-3'；
The gene order of PCV2-F2 is 5'-AGGGCCAGAATTCAACCTTT-3',
The gene order of PCV2-R2 is 5'-TCCCACAGAGTCTCAGTAGA-3'；
The gene order of PCV2-F3 is 5'-AGGGCCAGAATTCAACCTTAACCTTC-3',
The gene order of PCV2-R3 is 5'-GTTGAATTCTGGCCCTGCTCCCCGAT-3'.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105920595A (en) * | 2016-06-23 | 2016-09-07 | 江苏省农业科学院 | Porcine circovirus type 2 inactivated vaccine and preparation method thereof |
| CN108144057A (en) * | 2018-01-10 | 2018-06-12 | 杭州洪桥中科基因技术有限公司 | A kind of vaccine diluent and preparation method thereof |
| CN112618710A (en) * | 2019-09-24 | 2021-04-09 | 华南理工大学 | Phytoglycogen pig oral vaccination nano adjuvant and preparation method and application thereof |
| CN112679606A (en) * | 2020-12-31 | 2021-04-20 | 天津瑞普生物技术股份有限公司 | Hyperimmunity serum of erysipelothrix rhusiopathiae and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105920595A (en) * | 2016-06-23 | 2016-09-07 | 江苏省农业科学院 | Porcine circovirus type 2 inactivated vaccine and preparation method thereof |
| CN105920595B (en) * | 2016-06-23 | 2019-09-17 | 江苏省农业科学院 | Porcine circovirus 2 type inactivated vaccine and preparation method thereof |
| CN108144057A (en) * | 2018-01-10 | 2018-06-12 | 杭州洪桥中科基因技术有限公司 | A kind of vaccine diluent and preparation method thereof |
| CN112618710A (en) * | 2019-09-24 | 2021-04-09 | 华南理工大学 | Phytoglycogen pig oral vaccination nano adjuvant and preparation method and application thereof |
| CN112618710B (en) * | 2019-09-24 | 2023-09-29 | 华南理工大学 | Plant glycogen pig oral vaccination nanometer adjuvant and preparation method and application thereof |
| CN112679606A (en) * | 2020-12-31 | 2021-04-20 | 天津瑞普生物技术股份有限公司 | Hyperimmunity serum of erysipelothrix rhusiopathiae and preparation method thereof |
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