CN105833267A - Preparation method of novel adjuvant for haemophilus parasuis disease inactivated vaccines - Google Patents
Preparation method of novel adjuvant for haemophilus parasuis disease inactivated vaccines Download PDFInfo
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- 229940031551 inactivated vaccine Drugs 0.000 title claims abstract description 34
- 241000606807 Glaesserella parasuis Species 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 201000010099 disease Diseases 0.000 title abstract 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title abstract 2
- 235000008100 Ginkgo biloba Nutrition 0.000 claims abstract description 40
- 241000894006 Bacteria Species 0.000 claims abstract description 19
- 210000002966 serum Anatomy 0.000 claims abstract description 16
- 239000000243 solution Substances 0.000 claims abstract description 16
- 229960005486 vaccine Drugs 0.000 claims abstract description 15
- 244000194101 Ginkgo biloba Species 0.000 claims abstract description 11
- 238000000605 extraction Methods 0.000 claims abstract description 5
- 239000000427 antigen Substances 0.000 claims abstract description 4
- 102000036639 antigens Human genes 0.000 claims abstract description 4
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- 239000002574 poison Substances 0.000 claims description 31
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- 230000036039 immunity Effects 0.000 claims description 16
- 238000012360 testing method Methods 0.000 claims description 9
- 238000002347 injection Methods 0.000 claims description 8
- 239000007924 injection Substances 0.000 claims description 8
- 230000003053 immunization Effects 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 6
- 238000001514 detection method Methods 0.000 claims description 5
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- 241000193830 Bacillus <bacterium> Species 0.000 claims description 4
- 210000001744 T-lymphocyte Anatomy 0.000 claims description 4
- 238000002649 immunization Methods 0.000 claims description 4
- -1 terpene lactone Chemical class 0.000 claims description 4
- 230000010100 anticoagulation Effects 0.000 claims description 3
- 238000010241 blood sampling Methods 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 235000007586 terpenes Nutrition 0.000 claims description 3
- 235000009827 Prunus armeniaca Nutrition 0.000 claims 1
- 244000018633 Prunus armeniaca Species 0.000 claims 1
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- 238000007710 freezing Methods 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 12
- 238000000034 method Methods 0.000 abstract description 4
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- 239000000126 substance Substances 0.000 abstract description 3
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- 231100000419 toxicity Toxicity 0.000 abstract description 2
- 230000001988 toxicity Effects 0.000 abstract description 2
- 239000009429 Ginkgo biloba extract Substances 0.000 abstract 1
- 229940068052 ginkgo biloba extract Drugs 0.000 abstract 1
- 235000020686 ginkgo biloba extract Nutrition 0.000 abstract 1
- 230000008975 immunomodulatory function Effects 0.000 abstract 1
- 239000011550 stock solution Substances 0.000 abstract 1
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- 206010016256 fatigue Diseases 0.000 description 4
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- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 230000002779 inactivation Effects 0.000 description 3
- 230000000630 rising effect Effects 0.000 description 3
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- 102000004127 Cytokines Human genes 0.000 description 2
- 241000218791 Ginkgoaceae Species 0.000 description 2
- 206010018691 Granuloma Diseases 0.000 description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 2
- 229910052782 aluminium Inorganic materials 0.000 description 2
- 239000004411 aluminium Substances 0.000 description 2
- MOLPUWBMSBJXER-YDGSQGCISA-N bilobalide Chemical compound O([C@H]1OC2=O)C(=O)[C@H](O)[C@@]11[C@@](C(C)(C)C)(O)C[C@H]3[C@@]21CC(=O)O3 MOLPUWBMSBJXER-YDGSQGCISA-N 0.000 description 2
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- 238000010255 intramuscular injection Methods 0.000 description 2
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- 210000005259 peripheral blood Anatomy 0.000 description 2
- 239000011886 peripheral blood Substances 0.000 description 2
- XOJVVFBFDXDTEG-UHFFFAOYSA-N pristane Chemical compound CC(C)CCCC(C)CCCC(C)CCCC(C)C XOJVVFBFDXDTEG-UHFFFAOYSA-N 0.000 description 2
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010029350 Neurotoxicity Diseases 0.000 description 1
- 230000024932 T cell mediated immunity Effects 0.000 description 1
- 210000000662 T-lymphocyte subset Anatomy 0.000 description 1
- 206010044221 Toxic encephalopathy Diseases 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- 159000000013 aluminium salts Chemical class 0.000 description 1
- 229910000329 aluminium sulfate Inorganic materials 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 230000005784 autoimmunity Effects 0.000 description 1
- 230000007969 cellular immunity Effects 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 229930182486 flavonoid glycoside Natural products 0.000 description 1
- 150000007955 flavonoid glycosides Chemical class 0.000 description 1
- 244000144992 flock Species 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 229930184727 ginkgolide Natural products 0.000 description 1
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- 229930182470 glycoside Natural products 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000007365 immunoregulation Effects 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
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- 230000007246 mechanism Effects 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000003340 mental effect Effects 0.000 description 1
- 230000007135 neurotoxicity Effects 0.000 description 1
- 231100000228 neurotoxicity Toxicity 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 230000002085 persistent effect Effects 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 231100000397 ulcer Toxicity 0.000 description 1
- 239000012646 vaccine adjuvant Substances 0.000 description 1
- 229940124931 vaccine adjuvant Drugs 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/02—Bacterial antigens
- A61K39/102—Pasteurellales, e.g. Actinobacillus, Pasteurella; Haemophilus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/52—Bacterial cells; Fungal cells; Protozoal cells
- A61K2039/521—Bacterial cells; Fungal cells; Protozoal cells inactivated (killed)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
- A61K2039/552—Veterinary vaccine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55588—Adjuvants of undefined constitution
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Mycology (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention relates to a preparation method for a novel adjuvant for haemophilus parasuis disease inactivated vaccines .The method comprises the following steps of 1, separating and extracting bacterial strains and preparing bacteria solution antigen; 2, preparing a vaccine stock solution; 3, preparing finished vaccines, wherein a ginkgo biloba extract is selected as an adjuvant, and ginkgo biloba polyvalent inactivity vaccines of different concentrations, components and contents are prepared and obtained .The effective components of polysaccharose substances of the adjuvant are prepared through separation and extraction of traditional Chinese herbal medicine ginkgo biloba plants and then combined with haemophilus parasuis polyvalent inactivity vaccines, and the haemophilus parasuis ginkgo biloba adjuvant inactivated vaccines are prepared .The inactivated vaccines have the good immune modulating function, can regulate lymphocyte and phagocyte activity, can regulate the cell factor level in body serum, and have the advantages of being natural, low in toxicity, free of drug residues and the like .
Description
Technical field
The present invention relates to the preparation method of a kind of new adjuvant of Haemophilus parasuis inactivated vaccine, belong to
Technical field in pharmaceutical preparation.
Background technology
Traditional vaccine is many with aluminium glue, mineral oil, propolis as vaccine adjuvant, and aluminium salt is in many situations
Under be not a good adjuvant, especially in inducing cellular immune response.When aluminium adjuvant leads to
Granuloma would generally be produced when crossing the path of injection rather than intramuscular injection in subcutaneous or skin.
Another one side effect is just the increase in the generation of IgE, allergy and possible neurotoxicity.
Mineral oil adjuvant, due to the shortcoming that can not be metabolized of existence, therefore can produce a series of side effect:
Inflammatory reaction, granuloma, ulcer and heating etc. including injection site.It addition, mineral oil with
And component such as pristane, hexadecane etc. therein can cause the autoimmunities such as adjuvant type arthritis
Reaction.Therefore, mineral oil emulsion adjuvant toxicity is bigger.Propolis has antibacterial, antiviral, anti-swollen
Knurl, anti-inflammatory, enhancing body's immunity and promote regeneration effect, using as adjuvant,
Effective cellular immunity can be excited, can overcome the conventional inactivated vaccine can not effective activated cell
The weakness of immunity.But method for extracting propolis is very big on the impact of its activity, different solvents and extraction
The component difference that method is extracted is the biggest.Propolis composition depends on source and flora simultaneously,
It is caused the biggest obstacle in medicinal application and quality standardization by the diversity of composition.This is just
Adding difficulty for studying its mechanism of action further, also the application for propolis adjuvant causes
Impact.Meanwhile, the immunoregulation effect of propolis presents a certain amount effect relationship, and little dose has immunity
Humidification, and heavy dose has immunosuppressive action.Therefore its effective as adjuvant is studied
The adjuvant proportioning of composition.Due to propolis complicated component, effects affecting activity is many, also has
Mechanism also needs to continue to verify.
Have based on this, propose the present invention.
Summary of the invention
For the above-mentioned technical problem of prior art, it is an object of the invention to provide a kind of secondary pig addicted to
The preparation method of the new adjuvant of blood bacillosis inactivated vaccine, divides from Chinese traditional herbs Ginkgoaceae plant
From, extract and prepare adjuvant active princlple polysaccharose substance, then go out with haemophilus parasuis infection multivalence
Live vaccine combines, and prepares haemophilus parasuis infection ginkgo leaf adjuvant inactivated vaccine, has very well
Immunoloregulation function, can regulate lymphocyte, phagocyte activity, regulation body serum in
Cytokine levels etc., have the advantages such as natural, low toxicity, drug residue free.
For reaching above-mentioned purpose, the present invention is achieved by the following technical solutions:
The preparation method of a kind of new adjuvant of Haemophilus parasuis inactivated vaccine, comprises the following steps:
(1) separation and Extraction bacterial strain prepare bacterium solution antigen；
(2) vaccinogen liquid is prepared；
(3) vaccine finished product is prepared: choosing ginkgo biloba p.e is adjuvant, prepares variable concentrations
The ginkgo leaf multivalent inactivated vaccine of component content.
In the immunizing dose of 2mL/ head, the amount of adjuvant is 2.5mL-12.5mL.
Described ginkgo biloba p.e stoste every ml 40mg Han extract, wherein composition containing ginkgo flavones
Glycosides 9.6mg and terpene lactone 2.4mg.
Described ginkgo leaf multivalent inactivated vaccine is noted by musculi colli at piglet 15 age in days respectively
Penetrating test group piglet after 2mL, head exempt from 21 days and carry out the most immune, immunization route and dosage are same
Head exempts from, haemophilus parasuis bacillus serum 4 type after two exempt from latter 14 days, to test group piglet
Carry out attacking poison with 5 types, meanwhile, before and after attacking poison, adopt anticoagulation respectively for T cell subgroup stream
Formula detects.
Described poison mode of attacking is lumbar injection bacterium solution 3ml/ head, and 4 types and 5 type viable bacteria contents are equal
It is 7.5 × 109CFU, each group respectively before immunity and two exempt from after the 7th, 14 days and after attacking poison the 7th,
Blood sampling in 14 days, 4 DEG C of 4000r/min are centrifuged 25min, and to separate-70 DEG C of freezen protective of blood standby.
This by adding in vaccine bacterium solution by ginkgo biloba p.e, effective by addition
Controlling to obtain inactivated vaccine, inactivated vaccine is through 2 immunity, it is possible to effective improve pig secondary addicted to
Blood bacillus Hps antibody titer, reaches to be effectively improved porcine cytokine IL-2, IL-4 level, adds
The piglet adding the immunity of ginkgo leaf adjuvant inactivated vaccine attacks poison one week after, piglet peripheral blood
CD3+CD4+CD8+ quantity significantly increases, it is achieved that the vaccine of intramuscular injection various dose all can be real
Its 100% protected effect that haemophilus parasuis infection serum 4 type and Serotype 5 are attacked existing, for
Pathogenic haemophilus parasuis infection provides new generation vaccine and immunization method.
Accompanying drawing explanation
Fig. 1 difference adjuvant inactivated vaccine Hps antibody titer；
Fig. 2 respectively organizes IL-2 level in piglet serum；
Fig. 3 respectively organizes IL-4 level in piglet serum；
Fig. 4 respectively organizes piglet periphery blood T lymphocyte Flow cytometry result.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is further illustrated, but the protection of the present invention
Scope is not limited to this.
The preparation method of the new adjuvant of Haemophilus parasuis inactivated vaccine of the present invention, including following step
Rapid:
(1) separation and Extraction bacterial strain prepare bacterium solution antigen；
(2) vaccinogen liquid is prepared；
(3) vaccine finished product is prepared: choosing ginkgo biloba p.e is adjuvant, prepares variable concentrations
The ginkgo leaf multivalent inactivated vaccine of component content.
In the immunizing dose of 2mL/ head, the amount of adjuvant is 2.5mL-12.5mL.
Ginkgo biloba p.e stoste every ml 40mg Han extract, wherein composition containing ginkgo flavonoid glycoside
9.6mg, terpene lactone 2.4mg (ginkgolides, Bilobalide).
Application Example
1, choosing ginkgo biloba p.e is the ginkgo leaf polyvalent inactivation that adjuvant is prepared as containing variable concentrations
Vaccine, adjuvant concentration proportioning see table 1, is injected by musculi colli at piglet 15 age in days respectively
After exempting from 21 days containing adjuvant polyvalent inactivation vaccine 2mL, head, test group piglet carries out second time immunity,
Immunization route and dosage are exempted from head, after two exempt from latter 14 days, bloodthirsty to the secondary pig of test group piglet
Bacillus bacillus serum 4,5 type carries out attacking poison, and attacking poison mode is lumbar injection bacterium solution 3ml/ head, 4,
5 type viable bacteria contents are 7.5 × 109CFU, sets up multivalence seedling and blank, control group simultaneously
Lumbar injection PBS, each group respectively before immunity and two exempt from after the 7th, 14 days and after attacking poison the 7th,
Blood sampling in 14 days, 4 DEG C of 4000r/min are centrifuged 25min, and to separate-70 DEG C of freezen protective of serum standby；
Meanwhile, before and after attacking poison, anticoagulation is adopted respectively for T cell subgroup flow cytometer detection.
Table 1 haemophilus parasuis polyvalent inactivation seedling Chinese Herbal Medicines Adjuvant concentration
2, variable concentrations adjuvant inactivated vaccine attacks poison result
To add the haemophilus parasuis inactivated vaccine immunity piglet of the ginkgo leaf adjuvant of three kinds of concentration
After, adjuvant test group piglet lumbar injection is compared with haemophilus parasuis serum 4 type and 5 of strong virus force
Type bacterium solution 3ml/ head, viable bacteria content is 7.5 × 109CFU, result is as follows:
Ginkgo leaf high concentration group serum 4 type bacterium solution occurs flocking together after attacking poison 6h, reposes, spirit not
The symptom such as shaking, attack dead 4 piglets after poison 9h, protective rate is only 20%；Ginkgo leaf high concentration
The symptoms such as group Serotype 5 bacterium solution occurs flocking together after attacking poison 4h, reposes, lassitude, attack poison
Dead 4 piglets after 33h, protective rate is only 20%；Concentration group serum 4 type bacterium solution in ginkgo leaf
Occur flocking together after attacking poison 16h, repose, the symptom such as lassitude, attack after poison 19h dead 4
Piglet, protective rate is only 20%；In ginkgo leaf, concentration group Serotype 5 bacterium solution goes out after attacking poison 23h
Now flock together, repose, the symptom such as lassitude, attack dead 2 piglets after poison 46h, protective rate
It is 60%；Ginkgo leaf low concentration group serum 4 type bacterium solution occurs flocking together after attacking poison 13h, reposes,
The symptoms such as lassitude, attack dead 1 piglet after poison 124h, and protective rate is 80%；Ginkgo leaf
Low concentration group Serotype 5 bacterium solution occurs reposing after attacking poison 5h, and mental restoration after 3 days is all deposited
Living, protective rate is 100%.
Result of the test shows in ginkgo leaf additive inactivated vaccine, with 2mg/ml low concentration addition
The inactivated vaccine made, immune effect is preferable, the protective rate to haemophilus parasuis infection serum 4 type
Up to 80%, to the protective rate of haemophilus parasuis infection Serotype 5 up to 100%, it is shown in Table 2.
Table 2 variable concentrations Chinese Herbal Medicines Adjuvant vaccine haemophilus parasuis attacks poison result
3, different adjuvant inactivated vaccine Hps antibody titer detections
Add ginkgo leaf adjuvant inactivated vaccine Hps antibody titer testing result, ginkgo leaf adjuvant group
Hps antibody all raises, and wherein two exempt from latter 1 week, and ginkgo leaf adjuvant group is slightly above control group, and two
Exempting from latter 2 weeks, ginkgo leaf adjuvant group is significantly higher than control group, and two exempt from latter 2 weeks, ginkgo leaf adjuvant
Group antibody horizontal is the most in rising trend, and control group is without significant change, as seen Fig. 1.
4, porcine cytokine IL-2, IL-4 horizontal detection
(1) each adjuvant group different phase IL-2 level change general trend is: exempt to two before immunity
Latter 1 week on a declining curve, the most in rising trend, within 2 weeks, reaches peak after exempting to two；Attack poison
Rear IL-2 persistent levels declines.Ginkgo leaf low concentration group exempt from two after 1 thoughtful attack poison after 1 week water
Flat higher than control group, after wherein ginkgo leaf low concentration group is exempted from two 1 week, two exempted from 2 weeks and attack afterwards
After poison, 1 week interval level is higher than control group, such as Fig. 2.
(2) general trend of each adjuvant group different phase IL4 level is: to after immunity before immunity
1 week in rising trend, the most on a declining curve, within 2 weeks, reaches minimum after exempting to two, and two exempt from
Afterwards to attack after poison 2 weeks in slow ascendant trend, after ginkgo leaf low concentration group is exempted to two before immunity
Interval IL4 level is higher than multivalence control group, wherein higher with ginkgo leaf low concentration group level, as
Fig. 3.
5, the detection of T lymphocyte subsets
The piglet adding the immunity of ginkgo leaf adjuvant inactivated vaccine attacks poison one week after, piglet peripheral blood
CD3+CD4+CD8+ quantity, ginkgo leaf adjuvant group is significantly higher than control group, such as Fig. 4.
As fully visible, the present invention separates, extracts and prepares from Chinese traditional herbs Ginkgoaceae plant
Adjuvant active princlple polysaccharose substance, then combine with haemophilus parasuis infection multivalent inactivated vaccine,
Prepare haemophilus parasuis infection ginkgo leaf adjuvant inactivated vaccine, there is good immunoloregulation function,
Cytokine levels etc. in lymphocyte, phagocyte activity, regulation body serum can be regulated,
There is the advantages such as natural, low toxicity, drug residue free.
Above-described embodiment is only used for illustrating the inventive concept of the present invention, rather than weighs the present invention
The restriction of profit protection, all changes utilizing this design that the present invention carries out unsubstantiality, all should fall
Enter protection scope of the present invention.
Claims (5)
1. the preparation method of the new adjuvant of Haemophilus parasuis inactivated vaccine, it is characterised in that include
Following steps:
(1) separation and Extraction bacterial strain prepare bacterium solution antigen；
(2) vaccinogen liquid is prepared；
(3) vaccine finished product is prepared: choosing ginkgo biloba p.e is adjuvant, prepares variable concentrations composition
The ginkgo leaf multivalent inactivated vaccine of content.
2. the preparation method of the new adjuvant of Haemophilus parasuis inactivated vaccine as claimed in claim 1, its
Being characterised by: in the immunizing dose of 2mL/ head, the amount of adjuvant is 2.5mL-12.5mL.
3. the preparation method of the new adjuvant of Haemophilus parasuis inactivated vaccine as claimed in claim 1, its
It is characterised by: described ginkgo biloba p.e stoste every milliliter 40mg Han extract, wherein argentiferous
Apricot yellow ketoside 9.6mg and terpene lactone 2.4mg.
4. the new adjuvant of Haemophilus parasuis inactivated vaccine as described in claim 1-3 any claim
Preparation method, it is characterised in that: described ginkgo leaf multivalent inactivated vaccine is respectively piglet 15
After age in days is exempted from 21 days by musculi colli injection 2mL, head, test group piglet carries out second time immunity,
Immunization route and dosage are exempted from head, after two exempt from latter 14 days, bloodthirsty to the secondary pig of test group piglet
Bacillus bacillus serum 4 type and 5 types carry out attacking poison, meanwhile, adopt anticoagulation respectively before and after attacking poison
For T cell subgroup flow cytometer detection.
5. the preparation method of the new adjuvant of Haemophilus parasuis inactivated vaccine as claimed in claim 4, its
It is characterised by: described poison mode of attacking is lumbar injection bacterium solution 3ml/ head, 4 types and 5 type viable bacterias
Content is 7.5 × 109CFU, each group respectively before immunity and two exempt from after the 7th, 14 days and attack
Blood sampling in 7th, 14 days after poison, 4 DEG C of 4000r/min are centrifuged 25min and separate-70 DEG C of freezings of serum
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Cited By (5)
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| CN110339353A (en) * | 2019-06-18 | 2019-10-18 | 龙岩学院 | Application of the red clover as haemophilus parasuis vaccine adjuvant |
| CN110812474A (en) * | 2019-11-14 | 2020-02-21 | 山东滨州沃华生物工程有限公司 | Triple inactivated vaccine for porcine circovirus type 2, mycoplasma hyopneumoniae and haemophilus parasuis 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 |
| CN113730563A (en) * | 2021-09-15 | 2021-12-03 | 龙岩学院 | Haemophilus parasuis adjuvant vaccine and preparation method thereof |
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- 2016-05-04 CN CN201610289531.6A patent/CN105833267A/en active Pending
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| 田启超: "口蹄疫疫苗抗原加中药成分对微血管自律运动振幅的影响", 《万方数据》 * |
| 罗添添 等: "副猪嗜血杆菌灭活疫苗佐剂的筛选", 《四川畜牧兽医》 * |
| 谢培山: "银杏叶标准提取物EGb761及银杏叶制剂的质量评价（待续）", 《中国中药杂志》 * |
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| CN110339353A (en) * | 2019-06-18 | 2019-10-18 | 龙岩学院 | Application of the red clover as haemophilus parasuis vaccine adjuvant |
| 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 |
| CN110812474A (en) * | 2019-11-14 | 2020-02-21 | 山东滨州沃华生物工程有限公司 | Triple inactivated vaccine for porcine circovirus type 2, mycoplasma hyopneumoniae and haemophilus parasuis and preparation method 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 |
| CN113730563A (en) * | 2021-09-15 | 2021-12-03 | 龙岩学院 | Haemophilus parasuis adjuvant vaccine and preparation method thereof |
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