CN108588034A - A kind of rabbit hemorrhagic disease virus of variation and its application in preparing inactivated vaccine - Google Patents
A kind of rabbit hemorrhagic disease virus of variation and its application in preparing inactivated vaccine Download PDFInfo
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Abstract
Application the present invention provides a kind of rabbit hemorrhagic disease virus of variation and its in preparing inactivated vaccine, the strain are deposited in China typical culture collection center, SH17 plants of preservation entitled rabbit hemorrhagic disease virus variant, and deposit number is：CCTCC NO:V201806.The rabbit hemorrhagic disease virus sequence alignment that the strain is detached with the current country differs greatly, wherein the homology of major antigen gene only 80%.And the inclined young age of virus strain infection rabbit, current vaccine used for virus hemorrhagic disease of rabbit are not effectively protected effect to the strain.Therefore present invention firstly provides SH17 plants with rabbit hemorrhagic disease virus variant to prepare inactivated oil-emulsion vaccine, the vaccine has good immune effect, to the Small side effects of rabbit only are immunized, and the vaccine antigen prepared is good, it is safe, to external environment without any harmful effect, it is easy to pass through safety evaluation.
Description
Technical field
The present invention relates to animal doctor's infectious disease technical fields, specifically, being related to a kind of rabbit hemorrhagic disease disease of variation
Poison and its application in preparing inactivated vaccine.
Background technology
Rabbit hemorrhagic disease is acute, Septic blood infectious disease, which has high incidence and case fatality rate, be commonly called as
" rabbit pest ".The disease incubation period is short, only 1~3 day, and oxyhepatitis, spleen enlargement, Pulmonary hemorrhage, annulus trachealis is mainly presented in sick rabbit
There are hepatic necrosis, the typical cytopathics such as substantial viscera oedema, diffusivity bleeding in shape bleeding when serious.1984, the disease was for the first time
In Jiangsu Province, China large-scale outbreak, the whole nation is then spread to rapidly, and more than lethal 100,004,000 ten thousand rabbit are equal all over the world later
There is generation, is formed in global range and propagated.The normal Outbreak of this disease brings significant damage to the cultivation of rabbit industry, is to advise at present
One of the Infectious Diseases in modelling warren.In recent years, rabbit infects the inclined young age of rabbit hemorrhagic disease, and commercialized vaccine was immunized
Rabbit kind often there is the case where immuning failure.
Rabbit hemorrhagic disease virus (Rabbit hemorrhagic disease virus, RHDV) belongs to calicivirus
Section.This virus is not aggregated the red blood cell of horse, ox, sheep, dog, pig, chicken, duck, rabbit, rat, cavy and hamster, is only capable of agglutination people's
" O " type red blood cell, and do not influenced in a certain range by temperature, pH value, organic solvent and certain inorganic ions, the external world is supported
Drag is stronger, but can be inhibited by RHDV antiserum specificity.The virus is widely distributed in sick rabbit internal organs and body fluid, wherein with
Content highest in liver,spleen,kidney, lung and blood.The virus cannot still cultivate breeding in various primary or passage cell at present.
The best preventive measure at present is exactly that adjuvant Attenuated vaccine is taken to be immunized.Immune effect after addition adjuvant
It significantly improves, and length of holding time.Such as Zhuan Guohong uses subcutaneous injection method immunity inoculation RHDV oil emulsion vaccines, rabbit to exempt from
Epidemic disease generates the antibody level that antibody pole is significantly higher than Attenuated vaccine inoculation rabbit after being inoculated with 50 days, immune protective rate is high.It opens small
Fly to wait and rabbit pest aluminum hydroxide adjuvant vaccine and conventional organization inactivated vaccine are subjected to contrast test discovery, immunity inoculation aluminium hydroxide assistant
Agent seedling rabbit immunity inoculation after 60~90 days HI antibody titers reach 10log2More than, the vaccine holding time rose to 12 by 8 months
A month, stability was good, middle use easy to produce.
Invention content
The technical problem to be solved by the present invention is to provide a kind of rabbit hemorrhagic disease virus of variation and its go out in preparation
Application in live vaccine.The present invention is prepared for a kind of rabbit hemorrhagic disease inactivated oil-emulsion vaccine (SH17 using the variant
Strain), the prevalence of rabbit hemorrhagic disease caused by prevent the infection of RHDV variants.The vaccine Small side effects, and with preferable
Immunological safety.
The purpose of the present invention is what is be achieved through the following technical solutions：
In a first aspect, the present invention provides a kind of rabbit hemorrhagic disease virus, which is deposited in Chinese Typical Representative culture
Object collection, SH17 plants of preservation entitled rabbit hemorrhagic disease virus variant, deposit number are：CCTCC NO:
V201806。
SH17 plants of rabbit hemorrhagic disease virus variant in the present invention is inventor from the doubtful rabbit of Zhejiang farm
The natural sense contamination detached in the rabbit that dies of illness of viral hemorrhagic disease, is identified from specificity, virulence, immunogenicity etc.,
The strain that the virulence that filters out is strong, immunogenicity is good.
Second aspect, the present invention provides a kind of separation methods of rabbit hemorrhagic disease virus, include the following steps：
A, aseptic collection is died of illness the liver of rabbit, and toxic liver organization is ground, after centrifugation, takes supernatant, and is filtered and to be obtained filter
Liquid；
B, the filtrate for preparing step A is inoculated with the susceptible nonimmune rabbit of 45~55 ages in days, morbidity in 12~60h after acquisition inoculation
Dead rabbit has the liver organization of typical pathologic variation, passes through susceptible nonimmune rabbit continuous passage with toxic liver organization suspension
It cultivates, screens up to SH17 plants of rabbit hemorrhagic disease virus variant.
Preferably, in step A, the grinding is specially to be ground using the sterile saline that 5 times of amounts are added；It is described
Centrifugal condition is：4000-10000r/min centrifuges 10-30min；A diameter of 0.45 μm of the filter opening of the filter.
Preferably, in step B, the inoculum concentration of the filtrate is every leg muscle injection 1mL of susceptible nonimmune rabbit.
The third aspect, the present invention provides a kind of rabbit hemorrhagic disease virus according to claim 1 to prepare
Application in rabbit hemorrhagic disease inactivated oil-emulsion vaccine.
Fourth aspect, the present invention provides a kind of preparation method of rabbit hemorrhagic disease inactivated oil-emulsion vaccine, features
It is, includes the following steps：
S1, inoculation 45~55 malicious as kind using SH17 plants of rabbit hemorrhagic disease virus variant described in claim 1
The susceptible nonimmune rabbit of age in days, the liver of the dead rabbit of the interior morbidities of 12~60h after aseptic collection inoculation, and reject the connective group of liver
It knits；
S2, the liver for rejecting connective tissue is fully ground, is prepared into homogenate；Freeze thawing will be homogenized, be then added it is dual anti-, from
The heart takes supernatant, and shaking table inactivates after formaldehyde is added, and obtains inactivation tissue suspension；
S3, the inactivation tissue suspension obtained in step S2 is mixed with white-oil adjuvant, 4 DEG C save backup.
Preferably, in step S2, described is specially to be ground using the sterile saline that 5-10 times of volume is added；Institute
It is 3 times to state number of freezing and thawing.
Preferably, in step S2, it is described it is dual anti-be penicillin and streptomysin, the addition of penicillin and streptomysin is respectively
1000IU/mL；The centrifugal condition is：4000r/min centrifuges 30min.
Preferably, in step S2, final concentration of the 0.4% of the formaldehyde；The temperature of the shaking table inactivation is 37 DEG C, inactivates
Time is 48-72h.
5th aspect, the present invention provides a kind of rabbit hemorrhagic disease oil breast inactivation epidemic diseases prepared according to preceding method
Seedling.
Compared with prior art, the present invention has following advantageous effect：
SH17 plants of the rabbit hemorrhagic disease virus variant that the present invention is obtained, the rabbit viral detached with the current country
Haemorrhagic virus sequence alignment differs greatly, wherein the homology of major antigen gene only 80%.And the virus strain infection rabbit is partially young
Age, current vaccine used for virus hemorrhagic disease of rabbit are not effectively protected effect to the strain.
SH17 plants of the rabbit hemorrhagic disease virus variant that the present invention is obtained can stablize passage in rabbit body, according to
Inactivated oil-emulsion vaccine prepared by the strain has good protectiveness for the variant.Rabbit viral prepared by the present invention goes out
Mass formed by blood stasis inactivated oil-emulsion vaccine uses mature and stable production method, simple for process, vaccine antigen that is easy to operate, and preparing
It is good, it is safe, it is easy to pass through safety evaluation.
Description of the drawings
Upon reading the detailed description of non-limiting embodiments with reference to the following drawings, other feature of the invention,
Objects and advantages will become more apparent upon：
Fig. 1 is SH17 plants of the present invention and the Phylogenetic tree of RHDV is analyzed；
Fig. 2 is the RT-PCR amplifications of embodiment 1；Wherein：M is DL2000Marker；1 is pathological material of disease tissue；2 be the moon
Property control；
Fig. 3 is that the dissect of vaccine immunity potency test in embodiment 3 observes result；Wherein：Fig. 3 A are the tracheae of test group；
Fig. 3 B are the lungs of test group；Fig. 3 C are the liver of test group；Fig. 3 D are the tracheae of control group；Fig. 3 E are the lungs of control group；
Fig. 3 F are the liver of control group；
Fig. 4 is that the dissect of vaccine immunity potency test in embodiment 3 observes result；Wherein：The tracheae that Fig. 4 A are first group；
The lungs that Fig. 4 B are first group；The liver that Fig. 4 C are first group；The tracheae that Fig. 4 D are second group；The lungs that Fig. 4 E are second group；
The liver that Fig. 4 F are second group.
Specific implementation mode
With reference to specific embodiment, the present invention is described in detail.Following embodiment will be helpful to the technology of this field
Personnel further understand the present invention, but the invention is not limited in any way.It should be pointed out that the ordinary skill of this field
For personnel, without departing from the inventive concept of the premise, several changes and improvements can also be made.These belong to the present invention
Protection domain.
Rabbit hemorrhagic disease virus variant in the present invention is that inventor goes out from the doubtful rabbit viral in Zhejiang warren
The natural infection strain detached in the rabbit that dies of illness of mass formed by blood stasis.The variant of this separation and the antigenic specificity of traditional strain are larger, this
May be the reason of traditional rabbit hemorrhagic disease virus strain inactivated vaccine cannot protect rabbit kind to resist variant completely.
SH17 plants of rabbit hemorrhagic disease virus involved in the present invention is variant, with the tradition reported before the country
RHDV is compared, and sequence difference is larger between them, the relatively low (table of amino acid identity of major antigen gene (capsid protein)
1), phylogenetic analysis result is also shown, it and traditional RHDV epidemic strains genetic distance farther out, and are not located at same evolve and divide
Branch, but form an individual branches (Fig. 1).Illustrate that some tradition RHDV in SH17 and China is in different topology group, it was demonstrated that
SH17 plants are one plant of RHDV separation strains to make a variation, and test result further proves, existing commercialization rabbit hemorrhagic disease epidemic disease
Seedling cannot protect the rabbit (table 2) infected by RHDV variants completely.Therefore, the present invention does vaccine use for SH17 plants with RHDV variants
Kind poison is prepared for a kind of inactivated vaccine can be used for preventing the infection of RHDV variants.
The amino acid sequence homology comparison result of 1 SH17 plants of table and tradition RHDV strain capsid proteins
Embodiment 1
The separation identification that SH17 plants of rabbit hemorrhagic disease virus variant
The liver for acquiring the rabbit that dies of illness makees 5 times of dilution grindings with sterile saline, and 10000r/min centrifuges 10min, takes
Clearly, the filtering of 0.45 μm of filter is used in combination, obtains filtrate, be placed in -20 DEG C it is spare.
Specific primer, sense primer P1 (SEQ ID are designed according to the VP60 nucleic acid sequences of the RHDV announced on GenBank
No.1)：5 '-CGCACAGCGCCGCAAGGCGAAGC -3 ', downstream primer P2 (SEQ ID No.2)： 5’—
TCAGACATAAGAAAAGCCATTGG -3 ', amplification RHDV VP60 Gene Partial segments 1725bp.Before being extracted with Trizol methods
The RNA for stating the morbidity rabbit hepatic tissue of acquisition, is used in combination M-MLV reverse transcriptions at cDNA, it is used in combination to carry out RT-PCR amplifications as template,
Reaction condition is 95 DEG C of pre-degenerations 3min, 94 DEG C of denaturation 30s；55 DEG C of annealing 30s；72 DEG C extend 1min 45s；30 cycles, most
After extend 10min.After PCR, 1% agarose gel electrophoresis.As a result it is shown in one specific band of appearance at 1700bp
(Fig. 2).It is compared through BLAST, the VP60 genetic homologies for the RHDV that amplified fragments are detached with the country only have 80%, therefore tentatively sentence
The disconnected strain is RHDV variants.
Stability test and virulence identification
The susceptible nonimmune experiment rabbit of 4 health is taken, the tissue suspension handled well obtained by leg muscle injection abovementioned steps is (i.e.
Filtrate above-mentioned), every 1.0mL acquires the liver pathological material of disease of dead inoculation rabbit；The above test procedure of repetition, continuous 4 times.As a result
Show：The rabbit 100% for attacking poison is dead, and the death time in 18~48h, illustrate to obtain stable rabbit hemorrhagic disease sick
Poison, and SH17 plants are named as, it is deposited in China typical culture collection center, the entitled rabbit hemorrhagic disease of preservation
SH17 plants of virus variant, deposit number are：CCTCC NO:V201806 (preservation dates：On January 24th, 2018, depositary institution
Address：Hubei China province wuchang, wuhan Luo Jia Shan Wuhan University).It is cyclic annular that visible tracheae is dissected and observed in the bleeding of dead rabbit nostril
Bleeding, flush, liver bleeding is serious, illustrates that the separation strains have very strong pathogenicity, is velogen strain.
The secondary culture that SH17 plants of rabbit hemorrhagic disease virus variant
SH17 plants of rabbit hemorrhagic disease virus variant after preservation is made into 5 times of dilutions, inoculation with sterile saline
In the susceptible nonimmune rabbit of health of 45~55 ages in days.Every leg muscle injects 1mL.It is dead in 12~60h after aseptic collection inoculation
The rabbit kind died, aseptic collection has the liver organization of typical pathologic variation, and the connective tissue of rejecting thereon is as seed culture of viruses.
Using the SH17 plants of inoculations of strain for passing for 4~10 generations, the hepatic pathology tissue for the dead rabbit for rejecting connective tissue is added
The ratio of physiological saline grinding filtering, hepatic pathology tissue and sterile saline is 1:5(W/V).Penicillin, streptomysin is added
Each 1000IU/mL, 4000r/min centrifuge 30min, and Aspirate supernatant preserves in -20 DEG C.
It is prepared by 2 rabbit hemorrhagic disease tissue suspension of embodiment
SH17 plants of rabbit hemorrhagic disease virus variant after preservation (is obtained i.e. in embodiment 1 through secondary culture
Supernatant) with physiological saline make 5 times of dilutions, it is inoculated in the susceptible nonimmune rabbit of health of 45~55 ages in days.Every leg muscle note
Penetrate 1mL.Rabbit kind dead in 12~60h after aseptic collection inoculation, aseptic collection have the liver organization of typical pathologic variation, and
Connective tissue thereon is rejected as seed culture of viruses.
It is seeded in the susceptible nonimmune rabbit of health after seed culture of viruses above-mentioned is passed 4-10 generations, 12 after then inorganic acquisition is inoculated with~
The liver for the dead rabbit for rejecting connective tissue is added physiological saline grinding filtering, hepatic pathology by the liver of dead rabbit in 60h
The ratio of tissue and sterile saline is 1:5 (W/V), are prepared into homogenate.It is each that penicillin, streptomysin is added in freeze thawing 3 times
1000IU/mL, 4000r/min centrifuge 30min, take supernatant up to tissue suspension, and first is added according to the 0.4% of tissue suspension total amount
Aldehyde solution (final concentration 0.4%) is inactivated, and inactivation condition is：48~72h is inactivated in 37 DEG C of shaking tables.Nothing is carried out after inactivation
Bacterium is examined, inactivation is examined.The result is that without bacterial growth, and immune rabbit does not occur death.It is outstanding thus to obtain inactivation tissue
Liquid.
3 vaccine evaluation of embodiment is tested
The preparation of inactivated vaccine
4 generation of biography of seed culture of viruses will be used in embodiment 2SH17 plants prepare inactivation tissue suspension and press 1 with white-oil adjuvant:1 ratio
Mixing, obtains inactivated vaccine, is saved backup in 4 DEG C.
Vaccine safety is tested
15 SPF new zealand rabbits are bought, it is random to divide three groups, every group 5.First group of subcutaneous vaccination 1mL inactivation epidemic diseases above-mentioned
Seedling；Second group of subcutaneous overdose is inoculated with 2mL inactivated vaccines above-mentioned；Third group is control group, and 1mL sterile physiologicals are subcutaneously injected
Brine.Every group of experiment rabbit is raised in isolator, is observed continuously one week.As a result, being showed no adverse reaction, show the inactivated vaccine
Safety is good.
Vaccine immunity potency test
10 SPF new zealand rabbits are bought, are divided to two groups, every group 5 at random.Every hypodermic injection the 4th generation of 1.0mL of test group
The inactivated vaccine of SH17 plants of preparations, another 5 healthy new zealand rabbits are as a contrast.3 weeks after immune, two groups of new zealand rabbits are attacked
Poison, every leg muscle inject the 2 times of lethal dose virus liquids prepared in example 1.Every group of experiment rabbit is raised in isolator, sees
It examines 1 week, test group new zealand rabbit is all strong to live, and control group is all dead.Protective rate of the vaccine to rabbit hemorrhagic disease
100%, the control group death rate 100% (table 2).All experiment rabbit dissects are observed, it is seen that the internal organs of test group have no apparent disease
Become；The bleeding of tracheae ring-type, lungs and the liver bleeding of control group are serious (Fig. 3).It can be seen that rabbit hemorrhagic disease of the present invention
Inactivated oil-emulsion vaccine (variant) has good immune protective.
15 SPF new zealand rabbits are bought, it is random to divide three groups, every group 5.First group of every subcutaneous vaccination the 4th generation of 1.0mL
The inactivated vaccine of SH17 plants of preparations, second group of every commercially available rabbit hemorrhagic disease virus tissue inactivation seedling of inoculation 1.0mL, third
Group is used as blank control.3 weeks after immune, first group and second group of new zealand rabbit are carried out to attack poison, every leg muscle injects 2 times
Lethal dose virus liquid.Every group of experiment rabbit is raised in isolator, observes 1 week, and first group of new zealand rabbit is all strong to live, second group
All morbidities.Experiment rabbit dissect is observed, it is seen that first group of internal organs have no apparent lesion；Second group of tracheae ring-type bleeding,
Lungs and liver bleeding are serious, see Fig. 4.It can be seen that rabbit hemorrhagic disease inactivated oil-emulsion vaccine (variant) of the present invention is compared with city
The invasion of rabbit kind resistance rabbit hemorrhagic disease virus variant can be protected by selling rabbit hemorrhagic disease virus tissue inactivation seedling more.
2 SH17 plants of inactivated vaccine of table attacks poison protection result with commercial available vaccines
Specific embodiments of the present invention are described above.It is to be appreciated that the invention is not limited in above-mentioned
Particular implementation, those skilled in the art can make a variety of changes or change within the scope of the claims, this not shadow
Ring the substantive content of the present invention.In the absence of conflict, the feature in embodiments herein and embodiment can arbitrary phase
Mutually combination.
Sequence table
<110>China Agriculture Academe Shanghai Veterinary Institute（China Animal Health and Epidemiology Center Shanghai branch center）
<120>A kind of rabbit hemorrhagic disease virus of variation and its application in preparing inactivated vaccine
<130> DAG35763
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 23
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 1
cgcacagcgc cgcaaggcga agc 23
<210> 2
<211> 23
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 2
tcagacataa gaaaagccat tgg 23
Claims (10)
1. a kind of rabbit hemorrhagic disease virus, which is characterized in that the strain is deposited in China typical culture collection center,
SH17 plants of preservation entitled rabbit hemorrhagic disease virus variant, deposit number are：CCTCC NO:V201806.
2. a kind of separation method of rabbit hemorrhagic disease virus according to claim 1, which is characterized in that including following
Step：
A, aseptic collection is died of illness the liver of rabbit, and toxic liver organization is ground, after centrifugation, takes supernatant, and filter to get filtrate；
B, the filtrate for preparing step A is inoculated with the susceptible nonimmune rabbit of 45~55 ages in days, and morbidity is dead in 12~60h after acquisition inoculation
Rabbit has the liver organization of typical pathologic variation, is trained by susceptible nonimmune rabbit continuous passage with toxic liver organization suspension
It educates, screens up to SH17 plants of rabbit hemorrhagic disease virus variant.
3. the separation method of rabbit hemorrhagic disease virus according to claim 2, which is characterized in that described in step A
Grinding is specially to be ground using the sterile saline that 5 times of amounts are added；The centrifugal condition is：4000-10000r/min
Centrifuge 10-30min；A diameter of 0.45 μm of the filter opening of the filter.
4. the separation method of rabbit hemorrhagic disease virus according to claim 2, which is characterized in that described in step B
The inoculum concentration of filtrate is that every leg muscle of susceptible nonimmune rabbit injects 1mL.
5. a kind of application of rabbit hemorrhagic disease virus in preparing rabbit hemorrhagic disease inactivated oil-emulsion vaccine.
6. a kind of preparation method of rabbit hemorrhagic disease inactivated oil-emulsion vaccine, includes the following steps：
S1, using SH17 plants of rabbit hemorrhagic disease virus variant described in claim 1 as kind of a poison, 45~55 ages in days of inoculation are easy
Feel nonimmune rabbit, the liver of the dead rabbit of the interior morbidities of 12~60h after aseptic collection inoculation, and reject the connective tissue of liver；
S2, the liver for rejecting connective tissue is fully ground, is prepared into homogenate；It will be homogenized freeze thawing, dual anti-, centrifugation is then added,
Supernatant is taken, shaking table inactivates after formaldehyde is added, and obtains inactivation tissue suspension；
S3, the inactivation tissue suspension obtained in step S2 is mixed with white-oil adjuvant, 4 DEG C save backup.
7. the preparation method of rabbit hemorrhagic disease inactivated oil-emulsion vaccine according to claim 6, which is characterized in that step
In S2, described is specially to be ground using the sterile saline that 5-10 times of volume is added；The number of freezing and thawing is 3 times.
8. the preparation method of rabbit hemorrhagic disease inactivated oil-emulsion vaccine according to claim 6, which is characterized in that step
In S2, it is described it is dual anti-be penicillin and streptomysin, the addition of penicillin and streptomysin is respectively 1000IU/mL；The centrifugation item
Part is：4000r/min centrifuges 30min.
9. the preparation method of rabbit hemorrhagic disease inactivated oil-emulsion vaccine according to claim 6, which is characterized in that step
In S2, final concentration of the 0.4% of the formaldehyde；The temperature of shaking table inactivation is 37 DEG C, inactivation time 48-72h.
10. rabbit hemorrhagic disease inactivated oil-emulsion vaccine prepared by a kind of method according to claim 6.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810249237.1A CN108588034B (en) | 2018-03-22 | 2018-03-22 | Variant rabbit viral hemorrhagic disease virus and application thereof in preparation of inactivated vaccine |
Applications Claiming Priority (1)
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111686245A (en) * | 2020-06-29 | 2020-09-22 | 江苏省农业科学院 | Inactivated vaccine for rabbit hemorrhagic disease virus type 2 and preparation method thereof |
| CN113736752A (en) * | 2021-05-28 | 2021-12-03 | 中牧实业股份有限公司 | Rabbit viral hemorrhagic disease virus type 2 and application thereof in preparation of inactivated vaccine |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111686245A (en) * | 2020-06-29 | 2020-09-22 | 江苏省农业科学院 | Inactivated vaccine for rabbit hemorrhagic disease virus type 2 and preparation method thereof |
| CN111686245B (en) * | 2020-06-29 | 2023-03-14 | 江苏省农业科学院 | Inactivated vaccine for rabbit hemorrhagic disease virus type 2 and preparation method thereof |
| CN113736752A (en) * | 2021-05-28 | 2021-12-03 | 中牧实业股份有限公司 | Rabbit viral hemorrhagic disease virus type 2 and application thereof in preparation of inactivated vaccine |
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