CN105497888B - A kind of chicken Marek's disease heat resisting protective live vaccine and preparation method thereof - Google Patents
A kind of chicken Marek's disease heat resisting protective live vaccine and preparation method thereof Download PDFInfo
- Publication number
- CN105497888B CN105497888B CN201511000683.1A CN201511000683A CN105497888B CN 105497888 B CN105497888 B CN 105497888B CN 201511000683 A CN201511000683 A CN 201511000683A CN 105497888 B CN105497888 B CN 105497888B
- Authority
- CN
- China
- Prior art keywords
- heat resisting
- resisting protective
- disease
- live vaccine
- chicken
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
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
- A61K39/12—Viral antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/32—Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers, poly(meth)acrylates, or polyvinyl pyrrolidone
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/16011—Herpesviridae
- C12N2710/16311—Mardivirus, e.g. Gallid herpesvirus 2, Marek-like viruses, turkey HV
- C12N2710/16334—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Veterinary Medicine (AREA)
- General Health & Medical Sciences (AREA)
- Inorganic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Virology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention discloses a kind of chicken Marek's disease heat resisting protective live vaccines and preparation method thereof, belong to biotechnology.Chicken Marek's disease heat resisting protective live vaccine of the present invention by virus liquid with heat resisting protective is freeze-dried mixed forms；Its heat resisting protective prescription includes：Sucrose 8~10%, polyvinylpyrrolidone 2~4%, polylysine 3~6%, enzymolysis gelatin 1~2%, milk protein hydrolysate 2~6%, glutamine 0.4~1%, L-arginine 0.05~0.5%, glycerine 1~4%, Lubrol 2~5%；The present invention by the cold digestion of pancreatin, connect addition auxiliary liquid after poison and improve production efficiency, reduce cost；By adding heat resisting protective, reduces virus and freezing and damaged in process of vacuum drying, improve stability of the product in preservation and use；The method of the present invention prepare chicken Marek's disease heat resisting protective live vaccine, realize it is stored refrigerated in storage and transport process, prolonged shelf life to 2~8 DEG C preserve 36 months.
Description
Technical field
The present invention relates to a kind of heat resisting protective live vaccine and preparation method thereof, especially a kind of chicken Marek's disease is heat-resisting
Protective agent live vaccine and preparation method thereof, belongs to biotechnology.
Background technology
Marek's disease is a kind of lymphoproliferative neoplastic disease of chicken, is currently endanger poultry husbandry sound development three
There is higher hair in the chicken group of one of big main epidemic disease (Marek's disease, newcastle disease and Bursal Disease), infection
Sick rate and the death rate become a kind of worldwide disease.Its pathological manifestations is peripheral nerve lymphoid cell infiltration and increase, is caused
Limb (wing) is benumbed, and phymatoid lesion occurs at positions such as sexual gland, iris, various internal organs, muscle and skins.
Vaccine immunity is the important means of the prevention and control disease.There are two types of domestic common attenuated vaccines：One kind is for virus and carefully
The Marek's disease liquid nitrogen seedling that born of the same parents combine, its need Liquid nitrogen storage, transport and preservation condition harshness make the long-term preservation of vaccine
It is restricted with transport；Another kind takes off cellifugal herpes turkey virus freeze-dried live vaccine (HVT) for virus, preserves relatively
Easily, using extensive.
Currently, Marek's disease herpes turkey attenuated vaccine is produced using chicken embryo fibroblasts, freeze-drying is made mostly and lives
Vaccine.That there is production costs is higher for existing freeze dried vaccine production technology, preserves difficult and immune effect ideal not to the utmost etc. asks
Topic.Therefore, the Marek's disease live-vaccine that a kind of production cost is relatively low, strong suitable for large-scale production, defencive function is researched and developed
Heat resistant freeze drying protection technique is particularly important.
Invention content
The technical problem to be solved by the present invention is to overcome the defect of prior art, it is heat-resisting to provide a kind of chicken Marek's disease
Protective agent live vaccine and preparation method thereof, the chicken Marek's disease heat resisting protective live vaccine of preparation have that heat resistance is good, protects
The strong feature of protective function.
What the purpose of the present invention was achieved through the following technical solutions：
A kind of chicken Marek's disease heat resisting protective live vaccine is mixed with heat resisting protective by chicken Marek's disease virus
It is lyophilized；The heat resisting protective is made of the following component in terms of quality volumn concentration：Sucrose 8~10% gathers
Vinylpyrrolidone 2~4%, polylysine 3~6% digest gelatin 1~2%, milk protein hydrolysate 2~6%, glutamine
0.4~1%, L-arginine 0.05~0.5%, glycerine 1~4%, Lubrol 2~5%, surplus is water.
A kind of preparation method of chicken Marek's disease heat resisting protective live vaccine, includes the following steps：
A. prepared by cell：SPF chicken embryo fibroblasts are digested with EDTA- pancreatin, and cell growth medium is added, 36~37
It is cultivated 20~28 hours at a temperature of DEG C, until cellulation single layer；A concentration of quality volumn concentration of the EDTA- pancreatin is
1%~3%.
B. poison is connect：It is inoculated with chicken Marek's disease virus, the auxiliary culture solution of cell culture fluid volume 10% is added, by 100
~200 ten thousand/milliliter multiple cropping chicken embryo fibroblasts, 36~37 DEG C are continued culture 2~4.
C. harvest and freeze-drying：When cytopathy is up to 75%~85%, 90% cell culture fluid is discarded, rotates cell culture
Bottle makes cell all be detached from bottle wall, collects, low-speed centrifugal, takes sedimentation cell and weigh, and heat resisting protective is added, is homogenized
And ultrasonic treatment, quantitative separating and vacuum freezedrying, chicken Marek's disease heat resisting protective live vaccine is made.
The preparation method of above-mentioned chicken Marek's disease heat resisting protective live vaccine, the digestion temperature of EDTA- pancreatin in step a
It it is 2~8 DEG C, digestion time is 10~18 hours.
The preparation method of above-mentioned chicken Marek's disease heat resisting protective live vaccine, auxiliary culture solution is to contain water in step b
Solve the aqueous solution of lactoprotein, arginine and glutamine.
The preparation method of above-mentioned chicken Marek's disease heat resisting protective live vaccine, the addition of heat resisting protective in step c
100~300ml is added for every gram of sedimentation cell.
Vaccine of the present invention and preparation method thereof has the characteristics that：
(1) present invention adds surfactant Lubrol, amino acid polymer polylysine in heat resisting protective, and
It is most adapted to ratio with gelatin, polyvinylpyrrolidone etc. is digested, virus is reduced and is freezing and damaged in process of vacuum drying, and carry
High stability of the product in preservation and use.
(2) present invention uses the cold digestion SPF chicken embryos of high concentration EDTA- pancreatin, significantly improves cell yield and cell is lived
Property, combination cell multiple cropping technology and directly harvest sick cell effectively increase viral yield, significantly reduce cost, for production
High-titer vaccine lays the foundation.
(3) present invention is changed without cell culture fluid after connecing poison, but adds a small amount of auxiliary liquid, reduces production cost.
(4) the fowl heat resisting protective live vaccine invented, realizes stored refrigerated in vaccine storage and transport process, extends guarantor
The phase is deposited, being increased within 18 months 2~8 DEG C by original -15 DEG C or less preservations preserves 36 months.
Specific implementation mode
It is described further With reference to embodiment.
The preparation of 1 heat resisting protective of embodiment
The heat resisting protectives prescription raw materials such as sucrose, polyvinylpyrrolidone (K30) and enzymolysis gelatin are conventional reagent,
It is commercially available.
(1) heat resisting protective 1 is prepared
Prepare 1000ml heat resisting protectives：Weigh sucrose 100g, polyvinylpyrrolidone (K30) 40g, polylysine 60g,
Gelatin 10g, milk protein hydrolysate 20g, glutamine 4g, L-arginine 0.5g, glycerine 10ml, Lubrol 50g is digested to be added
In deionized water and constant volume is to 1000ml, heat resisting protective 1 is made, degerming is spare.
(2) heat resisting protective 2 is prepared
Prepare 1000ml heat resisting protectives：Weigh sucrose 80g, polyvinylpyrrolidone (K30) 30g, polylysine 30g,
Gelatin 20g, milk protein hydrolysate 60g, glutamine 10g, L-arginine 5g, glycerine 40ml, Lubrol 30g is digested to be added
In deionized water and constant volume is to 1000ml, heat resisting protective 2 is made, degerming is spare.
Auxiliary culture solution is the aqueous solution containing lactoalbumin hydrolysate, arginine and glutamine, lactoalbumin hydrolysate, arginine
With the content (m of glutamine:V) 50~150g/L, 0.70~2.10g/L and 1~3g/L are followed successively by.
The preparation of 2 chicken Marek's disease heat resisting protective live vaccine of embodiment
Marek isease turkey herpes virus (FC126 plants) seed culture of viruses is purchased from China Veterinery Drug Inspection Office.
(1) prepared by cell
10 age in days SPF chicken embryos are put into 2000ml screw sockets bottle (30 embryos/bottle), a concentration of 2% (m is added:V) EDTA- pancreases
Enzyme 45ml, digests SPF chicken embryos 16 hours under conditions of 4 DEG C, and filtered through gauze, filtrate centrifuges 10 minutes through 1500rpm, abandons
Clearly.60ml cell growth mediums are added into precipitation, cell suspension is made, and carries out cell count.It is with cell growth medium that cell is close
Degree is adjusted to 1,500,000/ml, accesses Tissue Culture Flask, and 36~37 DEG C are cultivated 24 hours, and fine and close cell monolayer is grown up to.This
The viable count that experiment digestion obtains fibrocyte is 3 × 108A/embryo, about the 2 of conventional method times.
(2) poison is connect
By volumn concentration (V：V) then auxiliary is added in cell culture fluid in 2% inoculation chicken Marek's disease virus
Culture solution, it is 10% (V of cell culture fluid volume percentage composition to assist the addition of culture solution：V), and it is multiple by 1,000,000/milliliter
Kind chicken embryo fibroblasts, 36~37 DEG C are continued to cultivate.Auxiliary culture solution is to contain lactoalbumin hydrolysate, arginine and glutamine
Aqueous solution, concentration is followed successively by 50g/L, 0.70g/L and 3g/L.
(3) harvest and freeze-drying
Culture was harvested to 72 hours when cytopathy is up to 80%, discarded 90% cell culture fluid, rotated Tissue Culture Flask,
So that cell is all detached from bottle wall, cell is collected in centrifugal bottle, 2000rpm is centrifuged 10 minutes, takes sedimentation cell, and weigh.
According to the amount that 200ml heat resisting protectives are added in every gram of sedimentation cell, it is separately added into the above-mentioned heat resisting protective 1 prepared and resistance to
Then thermal protecting agent 2 carries out homogenate and ultrasonic treatment, the cell liquid of cracking is finally carried out quantitative separating and freezing vacuum is done
It is dry, chicken Marek's disease heat resisting protective live vaccine 1 and chicken Marek's disease heat resisting protective live vaccine 2 is made.
According to No. 297 bulletins of the Ministry of Agriculture, (marek isease turkey herpes virus heat resisting protective live vaccine manufactures and inspection
Test Trial Regulation, similar heat-resisting seedling professional standard) method prepares similar heat-resisting seedling as a contrast 1；According to《The People's Republic of China (PRC)
Regulations》(2000 editions) (hereinafter referred to as " regulation ") method prepares conventional seedling as a contrast 2.
(4) vaccine stability measures
1. detecting stored refrigerated stability：Before freeze-drying, after freeze-drying 2~8 DEG C place 0,6,12,24 and at 36 months with
Viral level, 3 bottles/time are measured by sampling in machine.It the results are shown in Table 1
2. the stability of 37 DEG C of preservations of detection：Every batch of takes 20 bottles, grab sample survey when placing 0,5,10 and 15 day for 37 DEG C
Determine viral level, 3 bottles/time.It the results are shown in Table 2
The different storage life chicken Marek's disease virus of live vaccine content detection results (2~8 DEG C) of table 1
Unit：PFU/ plumages
| Group | Before freeze-drying | 0 month | June | December | 24 months | 36 months |
| Heat-resisting protective vaccinating agent 1 | 8000 | 7600 | 7500 | 7200 | 7000 | 6000 |
| Heat-resisting protective vaccinating agent 2 | 8000 | 7400 | 7400 | 7000 | 6800 | 6500 |
| Compare 1 (similar heat-resisting seedling) | 6000 | 5000 | 3900 | 3000 | 2600 | <2000 |
| Compare 2 (conventional seedlings) | 6000 | 4200 | 2800 | <1000 | <1000 | <1000 |
The different storage life chicken Marek's disease viral level testing results (37 DEG C) of table 2
Unit：PFU/ plumages
| Group | 0 day | 5 days | 10 days | 15 days |
| Heat-resisting protective vaccinating agent 1 | 7600 | 7000 | 6800 | 6000 |
| Heat-resisting protective vaccinating agent 2 | 7400 | 6900 | 6600 | 6200 |
| Compare 1 (similar heat-resisting seedling) | 5000 | 3200 | 2600 | <1000 |
| Compare 2 (conventional seedlings) | 4200 | <1000 | <1000 | <1000 |
As can be seen from the above results：The two groups of chicken Marek's disease live-vaccines prepared using the method for the present invention, 2~8
It is little that 36 months its viral level falls are preserved under the conditions of DEG C, viral level is far above chicken Marek's in " regulation " standard
Disease live-vaccine viral level answers >=requirements of 2000PFU/ plumages, storage life viral survival rate in 36 months time vaccines is up to
78%；It is preserved under the conditions of 2~8 DEG C of 1 (similar heat-resisting seedling) of control and control 2 (conventional seedling), viral level declines very fast, control 1
It is only 6 months that viral survival rate, which is only 52%, 2 storage lives of control, when storage life is 24 months, and 6 months viral survival rates are
67%.
It is little that 15 days its viral level falls are preserved under the conditions of 37 DEG C, viral level is provided far above " regulation ", is protected
It deposits phase viral survival rate in 15 days time vaccines and is up to 79%；Compare 1 (similar heat-resisting seedling), control 2 (conventional seedling) 37 DEG C of conditions
Lower preservation, viral level decline comparatively fast, and 1 storage life of control is 10 days, 2 storage lives of control were less than 5 days.
In addition, 30 pieces of this seedling SPF embryos dosage, is made 304 bottles of the heat-resisting seedling of 1000 plumages/bottle, 8000PFU/ before being lyophilized
Plumage, it is 8.1 × 10 that single embryo, which can produce virus quantity,7PFU is 4~10 times of (conventional method list embryo production virus quantities of conventional method
It is 0.8~2.0 × 107PFU)。
Claims (4)
1. a kind of chicken Marek's disease heat resisting protective live vaccine, which is characterized in that by chicken Marek's disease virus and heat-resisting guarantor
Agent is freeze-dried mixed forms for shield；The heat resisting protective is made of the component of following quality volumn concentration meter：Sucrose 8~
10%, polyvinylpyrrolidone 2~4%, polylysine 3~6% digests gelatin 1~2%, milk protein hydrolysate 2~6%, paddy
Glutamine 0.4~1%, L-arginine 0.05~0.5%, glycerine 1~4%, Lubrol 2~5%, surplus are water.
2. a kind of preparation method of chicken Marek's disease heat resisting protective live vaccine described in claim 1, which is characterized in that system
It is standby to carry out as follows：
A. prepared by cell：SPF chicken embryo fibroblasts are digested with EDTA- pancreatin, and cell growth medium is added, in 36~37 DEG C of temperature
The lower culture of degree 20~28 hours, until cellulation single layer；A concentration of quality volumn concentration of the EDTA- pancreatin is 1%
~3%；
B. poison is connect：Kind enters chicken Marek's disease virus, the auxiliary culture solution of cell culture fluid volume 10% is added, by 100~200
Ten thousand/milliliter multiple cropping chicken embryo fibroblasts, 36~37 DEG C are continued culture 2~4；
C. harvest and freeze-drying：When cytopathy is up to 75%~85%, 90% cell culture fluid is discarded, Tissue Culture Flask is rotated, makes
Cell is all detached from bottle wall, collects, low-speed centrifugal, takes sedimentation cell and weigh, and heat resisting protective is added, and carries out homogenate and ultrasound
Wave cracks, and chicken Marek's disease heat resisting protective live vaccine is made in quantitative separating and vacuum freezedrying；
It is the aqueous solution containing lactoalbumin hydrolysate, arginine and glutamine that culture solution is assisted in step b.
3. the preparation method of chicken Marek's disease heat resisting protective live vaccine according to claim 2, which is characterized in that step
The digestion temperature of EDTA- pancreatin is 2~8 DEG C in rapid a, and digestion time is 10~18 hours.
4. the preparation method of chicken Marek's disease heat resisting protective live vaccine according to claim 2, which is characterized in that step
The addition of heat resisting protective is that 100~300ml is added in every gram of sedimentation cell in rapid c.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201511000683.1A CN105497888B (en) | 2015-12-29 | 2015-12-29 | A kind of chicken Marek's disease heat resisting protective live vaccine and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201511000683.1A CN105497888B (en) | 2015-12-29 | 2015-12-29 | A kind of chicken Marek's disease heat resisting protective live vaccine and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105497888A CN105497888A (en) | 2016-04-20 |
| CN105497888B true CN105497888B (en) | 2018-07-27 |
Family
ID=55706435
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201511000683.1A Active CN105497888B (en) | 2015-12-29 | 2015-12-29 | A kind of chicken Marek's disease heat resisting protective live vaccine and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105497888B (en) |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1168503C (en) * | 2001-12-21 | 2004-09-29 | 卫广森 | Veterinarian virus kind biological product heat resisting freeze drying protective agent and its preparation technique |
| CN1222314C (en) * | 2001-12-21 | 2005-10-12 | 卫广森 | Heat resisting lyophilized protectant for chicken Marek's disense turkey herpes virus lyophilized vaccine and preparing process thereof |
| CN102000328B (en) * | 2010-11-23 | 2012-12-12 | 北京市兽医生物药品厂 | Method for manufacturing Marek's disease vaccine by utilizing cell factory |
| CN103127496B (en) * | 2011-11-30 | 2016-01-20 | 普莱柯生物工程股份有限公司 | Type III herpes turkey virus freeze dried vaccine |
| CN103977400B (en) * | 2014-05-29 | 2015-06-17 | 南京创启生物科技有限公司 | Method for producing marek disease live vaccine of chicken by using cell line |
-
2015
- 2015-12-29 CN CN201511000683.1A patent/CN105497888B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN105497888A (en) | 2016-04-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106237339B (en) | A kind of freeze dried vaccine heat resisting protective and its preparation method and application | |
| CN102220287B (en) | Avian infectious bronchitis cold adaptation attenuated vaccine strain and application thereof | |
| CN106119212B (en) | Fowl adenovirus strain, inactivated vaccine and preparation method | |
| CN111000993B (en) | Bivalent inactivated vaccine for duck viral hepatitis and duck reovirus disease and preparation method thereof | |
| CN105664150B (en) | A kind of newcastle disease virus, avian influenza virus and aviadenovirus triple inactivated vaccine | |
| CN108992674A (en) | A kind of heat resisting protective and its application | |
| CN105582533B (en) | Avian influenza virus and avian adenovirus bivalent inactivated vaccine | |
| CN106754762A (en) | A kind of antigen of encephalitis B live vaccine and preparation method and application | |
| CN110628726B (en) | Novel Muscovy duck adenovirus strain, bivalent inactivated vaccine and preparation method thereof | |
| CN110893235B (en) | Inactivated vaccine for crucian hematopoietic necrosis and preparation method thereof | |
| CN104306963B (en) | A kind of production method of pig erysipelas live vaccine | |
| CN104367996B (en) | A kind of method and its product that pseudorabies disease live-vaccine is produced with subculture cell source | |
| CN105561317B (en) | Recombinant pseudorabies virus vaccine heat-resisting lyophilized protecting agent and preparation method thereof | |
| CN111778217B (en) | I-group 4 type avian adenovirus SD-F strain, inactivated vaccine and preparation method | |
| CN101745106A (en) | Porcine parvnvirus living vaccine and preparation method thereof | |
| CN108815517A (en) | A kind of Duck plague live vaccine and preparation method thereof | |
| CN103143007B (en) | Method for preparing infectious bursal disease virus live vaccine by utilizing passage chicken embryo fibroblast | |
| CN111643659A (en) | Rabbit hemorrhagic disease virus baculovirus vector vaccine and preparation method thereof | |
| CN105497888B (en) | A kind of chicken Marek's disease heat resisting protective live vaccine and preparation method thereof | |
| CN105497904B (en) | A kind of pseudorabies virus vaccine heat-resisting lyophilized protecting agent and preparation method thereof | |
| CN109207436A (en) | One plant of 4 type aviadenovirus strain of I group and its application | |
| CN102000328A (en) | Method for manufacturing Marek's disease vaccine by utilizing cell factory | |
| CN105838641A (en) | Mycoplasma synoviae culture method | |
| CN112999343B (en) | Inactivated vaccine of goose astrovirus and preparation method thereof | |
| CN111411087B (en) | Cyprinus carpioides herpesvirus II type low virulent strain and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |