CN101905021B - Trigeminal live vaccine of canine distemper viruses, canine parvoviruses and Type I canine adenoviruses and preparation method thereof - Google Patents
Trigeminal live vaccine of canine distemper viruses, canine parvoviruses and Type I canine adenoviruses and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a trigeminal live vaccine of canine distemper viruses, canine parvoviruses and type I canine adenoviruses and a preparation method thereof. The trigeminal live vaccine comprises a canine distemper hypotoxicity vaccine strain with a microbiological collection number of CGMCC (China General Microbiological Culture Collection Center) No.3810, a canine parvovirus hypotoxicity vaccine strain with a microbiological collection number of CGMCC No.3841 and a type I canine adenovirus vaccine strain with a microbiological collection number of CGMCC No.3809. The trigeminal vaccine is prepared by adopting the canine distemper viruses, the canine parvoviruses and the type I canine adenoviruses which are wild viruses separated from Chinese sick canine and domesticated and attenuated as objects, and a freeze drying protective agent is also added during preparing. The trigeminal live vaccine has reliable safety, good immune effect and strong pertinence, is suitable for preventing diseases of Chinese sick canine, and can be used for preventing infectious diseases of canine distempers, the canine parvoviruses and canine infectious hepatitis.
Description
Technical field
The present invention relates to trigeminal live vaccine for animals, relate in particular to canine distemper, Canine Parvovirus and hepatitis infectiosa canis virus I type trigeminal live vaccine and preparation method thereof, belong to trigeminal live vaccine for animals field.
Background technology
Canine distemper, Canine Parvovirus and hepatitis infectiosa canis virus are currently dog is supported by China already to endanger 3 kinds of serious viral infectious.
Canine distemper is the acute infectious disease of animals such as a kind of dog class, acute, lethal process that mostly young animal is, and adults can be chronic persistent infection, faces that to examine with two-phase pattern of fever, mucosa mucositis, nervus centralis symptom and footpad swelling be principal character.This disease almost has been worldwide distribution since nineteen twenty-six comes to light, be currently dog industry, fur-bearing animal aquaculture and the conservation of wildlife are supported by China already to endanger one of maximum disease, often causes animals morbidities such as large quantities of dogs, ermine, Vulpes, and economic loss is heavy.In recent years; The multiple animal such as title Lagothrix and clasper order Phocidae that comprises all 8 sections of Carnivoras such as giant panda, lesser panda and lion, tiger, leopard, Artiodactyla Suidae, Primates all has the report of CD natural occurrence; Even the people also has the case of CDV infection; And the animal range of CDV natural infection also has the trend that constantly enlarges, its harm also increasing (Cai Baoxiang. livestock epidemiology (third edition). Beijing: the .2001 of Chinese agriculture publishing house, 347-351.).
(AppelM J, Scott FW, CarmichaelL E.Isolation and immunization studies of a canine parco-like virus from dogs with haemorrhagic enteritis.Vet Rec after Canine Parvovirus came to light in 1978 first; 1979,105 (8): 156-159.), primary disease is popular in all over the world; China is in nineteen eighty-two alleged occurrence Canine parvovirus infection (Liang Shizhe, canal river rose, Wei Xiren; Deng. the parvovirus granule that detects in the research I-diarrhoea dog feces of canine infectious enteritis. Shanghai animal and veterinary communication, 1982,2 (4): 172.); Next year Xu Hankun etc. has formally reported popular (Xu Hankun, Guo Baofa, the Jin Huai of primary disease; Deng. blood clotting and hemagglutination inhibition test break out answering in the Canine Parvovirus enteritis the dog crowd. Chinese poultry infectious disease, 1983 (4): 43-45.).Canine Parvovirus is cause of disease (the A fshar A..Canine Parvovirus infection-a review.Vet Bull that causes dog acute hemorrhagic gastroenteritis and pup acute myocarditis; 1981; 51:605-612.), the main harm pup, particularly 2～6 the monthly age dog.The morbidity of this disease is anxious, and the course of disease is short, and mortality rate is high, and infectiousness is strong, to experimental dog, and army dog, dog crowd such as police dog and pet dog all have very big danger.
I type hepatitis infectiosa canis virus comes to light in nineteen twenty-five first.1984, be separated to this virus first in China, confirmed also to have in China dog the infection (Yin Zhen of hepatitis infectiosa canis virus I type; Liu Jinghua. animal virology (second edition). Beijing: Science Press, 1997,757-762.) .I type hepatitis infectiosa canis virus can cause HCC (Webe J.DNA vaccine.BusinessWeek clinically; 1996,2: 6.), generation (Xia Xianzhu, the model spring water of The Fox and the Bear encephalitis; Hu Rongliang; Experimental immunization study Deng malicious YCA18 strain a little less than the .II type hepatitis infectiosa canis virus nature. Chinese veterinary drug magazine, 2001,35 (2): 1-4).
External is prevention and these several kinds of infectious disease of control, has developed multiple commodity with single Seedling and couplet Seedling; The report that agriculture and animal husbandry university of domestic existing PLA dog is succeeded in developing with 5-linked Seedling (rabies, canine distemper, dog parainfluenza, hepatitis infectiosa canis virus 2 type disease and parvovirus infection).
Join the vaccine that Seedling is meant that microorganism not of the same race or its metabolite are formed, the using dosage that joins Seedling is suitable with single Seedling, but its effectivenesss is identical with single Seedling, and joins Seedling and can simplify vaccine program, reduces the number of times of inoculating the animal emergency reaction, reaches the pin purposes of preventing more.The present invention successively separates obtaining Canine Parvovirus, hepatitis infectiosa canis virus I type Strain again on canine distemper list Seedling research basis, and has carried out the development of the sick I type of canine distemper, canine parvovirus disease and hepatitis infectiosa canis virus trigeminal live vaccine.
Summary of the invention
One of the object of the invention provides a kind of canine distemper, Canine Parvovirus and hepatitis infectiosa canis virus I type trigeminal live vaccine;
Two of the object of the invention provides a kind of method for preparing above-mentioned canine distemper, Canine Parvovirus and hepatitis infectiosa canis virus I type trigeminal live vaccine;
The present invention seeks to realize through following technical scheme:
A kind of canine distemper (Canine distemper virus), Canine Parvovirus (Canine parvovirus) and I type hepatitis infectiosa canis virus (Canine adenovirus) three attenuated live vaccines; Comprise: microbial preservation number is the canine distemper attenuated vaccine strain of CGMCC No.3810, and microbial preservation number is that Canine Parvovirus attenuated vaccine strain and the microbial preservation of CGMCC No.3841 number is the I type canine adenovirus attenuated vaccine strain of CGMCC No.3809.
Preferably, said canine distemper attenuated vaccine strain, malicious valency>=10 of Canine Parvovirus attenuated vaccine strain or I type canine adenovirus attenuated vaccine strain
4.5TCID
50/ head part.
The cultivation of canine distemper virus low virulent strain of the present invention (CDV-YBR): isolating canine distemper virus (CDV-YB strain) was passaged to for 70 generations through vaccinization CEF (CEF); Change afterwards at the Vero cell and uploaded for 20 generations, continue then to be uploaded to for 105 generations at CEF (CEF).In the process of going down to posterity, the titre of virus improves gradually, and strain more and more adapts to CEF and Vero cell.But virus lowers along with the increase of passage number the pathogenicity of dog gradually; Tests such as specificity through morphology evaluation, the check of pure property, virus and exogenous virus check, result verification the CDV-YB low virulent strain (CDV-YBR) cultivated have typical paramyxovirus particle characteristics, pure no exogenous virus pollutes.
The present invention submits the isolating canine distemper virus low virulent strain of institute (CDV-YBR) preservation of to patent accreditation body, and its microbial preservation number is: CGMCC No.3810; Classification called after: canine distemper virus; The preservation time is: on May 14th, 2010; Depositary institution is: China Committee for Culture Collection of Microorganisms common micro-organisms center; Preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica.
Through continuous synchronization inoculation CRFK passage to 110 generation, in the process of going down to posterity, the titre of virus improves gradually with the isolating Canine Parvovirus of the present invention (CPV-YN strain), and strain more and more adapts to the CRFK cell.In contrast, virus lowers along with the increase of passage number the pathogenicity of dog gradually; Through morphology identify, result of the tests such as the specificity of pure property check, virus and exogenous virus check have verified that the CPV-YN low virulent strain of cultivating (CPV-YNR) has typical parvovirus particle characteristics, pure no exogenous virus pollutes.The present invention with isolating CPV-YN low virulent strain (CPV-YNR) submit mechanism's preservation of patent approval to, its microbial preservation number is: CGMCC No.3841; Classification called after: Canine Parvovirus; The preservation time is: on May 14th, 2010; Depositary institution is: China Committee for Culture Collection of Microorganisms common micro-organisms center; Preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica.
The cultivation of the canine adenovirus attenuated strain CAV-HR of I type of the present invention: above-mentioned isolating hepatitis infectiosa canis virus (CAV-H strain) was passaged to for 120 generations through the vaccinization mdck cell, and in the process of going down to posterity, the titre of virus improves gradually, and strain more and more adapts to mdck cell.In contrast, virus lowers along with the increase of passage number the pathogenicity of dog gradually; Through morphology identify, result of the tests such as the specificity of pure property check, virus and exogenous virus check have verified that the CAV-H low virulent strain of cultivating (called after CAV-HR) has typical adenovirion characteristic, pure no exogenous virus pollutes.
The present invention with isolating CAV-H low virulent strain (called after CAV-HR) submit mechanism's preservation of patent approval to, its microbial preservation number is: CGMCC No.3809; Classification called after: hepatitis infectiosa canis virus; The preservation time is: on May 14th, 2010; Depositary institution is: China Committee for Culture Collection of Microorganisms common micro-organisms center; Preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica.
Another object of the present invention provides a kind of method of preparation above-mentioned canine distemper (Canine distemper virus), Canine Parvovirus (Canine parvovirus) and I type hepatitis infectiosa canis virus (Canine adenovirus) three attenuated live vaccines; Comprise: (1) cultivating microorganism preserving number respectively is the canine distemper attenuated vaccine strain of CGMCC No.3810; Microbial preservation number is that Canine Parvovirus attenuated vaccine strain and the microbial preservation of CGMCC No.3841 number is the I type canine adenovirus attenuated vaccine strain of CGMCC No.3809, obtains viral liquid;
(2) viral liquid is carried out titration of virus, mix, filter, add freeze drying protectant, lyophilizing promptly gets.
Wherein, the cultivation of said canine distemper attenuated vaccine strain comprises: select through aseptic, well-grown CEF monolayer that detection of mycoplasma is negative, remove culture fluid after, change to contain the cell maintenance medium of 2% seed culture of viruses, adjustment pH value 7.2～7.4 is put 33 ℃ of cultivations; Results when CPE reaches 70%.
The cultivation of said Canine Parvovirus attenuated vaccine strain comprises: seed culture of viruses is inoculated in through aseptic by 3% of cell maintenance medium amount synchronously, and well-grown CRFK monolayer that detection of mycoplasma is negative, the adjustment pH value is 7.2～7.4, puts 37 ℃ of cultivations; Treat that 70% cell CPE occurs and can gather in the crops.
The cultivation of said hepatitis infectiosa canis virus I type attenuated vaccine strain comprises: select through aseptic, well-grown MDCK monolayer that detection of mycoplasma is negative, remove culture fluid after, change to contain the cell maintenance medium of 2% seed culture of viruses, adjustment pH value 7.2～7.4; Put 37 ℃ of cultivations; Results when CPE reaches 70%.
The preparation of described freeze drying protectant comprises: (1) gelatin: 8g; Sucrose: 40g; Deionized water: 100ml; The adjustment pH value is 6.8～7.0; (2) 116 ℃ of sterilization 40min promptly get.
In the above-mentioned method for preparing, preferred, viral liquid is mixed in 9: 1 ratio with freeze drying protectant.
The present invention adopts separation, and canine distemper virus, Canine Parvovirus and hepatitis infectiosa canis virus I type malicious and that process is tamed a little less than causing are object from the open country of Chinese sick dog, prepare trigeminy vaccine together, and the adding freeze drying protectant are processed.This trigeminy vaccine safety is reliable, good immune effect and with strong points, is fit to the prevention of Chinese sick dog disease, can be used for preventing canine distemper, Canine Parvovirus, three kinds of infectious disease of HCC; The seedling craft science, program is workable, and economic benefit is considerable, is suitable for suitability for industrialized production.
The present invention with prepared canine distemper, parvovirus infection, adenopathy viral disease trigeminal live vaccine respectively 2～4 monthly age of immunity 15 of dogs, attack CDV-YB, CPV-YN, CAV-H poison by force after 21 days respectively, dosage is 200TCID
50/ dog.The result shows, 5 batches of vaccines all reach 4/5, equal 5/5 morbidity of contrast dog to the protective rate of three kinds of diseases.Therefore, trigeminal live vaccine vaccine of the present invention plays good effect to prevention canine distemper, parvovirus infection, adenovirus condition of disease.
Description of drawings
Fig. 1 CDV-YB 105 generation virion electromicroscopic photograph.
Fig. 2 CPV-YN 90 generation virion electromicroscopic photograph.
Fig. 3 CAV-H 110 generation virion electromicroscopic photograph.
The production process route figure of Fig. 4 trigeminal live vaccine of the present invention.
The specific embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage of the present invention and characteristics will be more clear along with description.But these embodiment only are exemplary, scope of the present invention are not constituted any restriction.It will be understood by those skilled in the art that and down can make amendment with form or replace without departing from the spirit and scope of the present invention, but these modifications and replacing all fall in protection scope of the present invention the details of technical scheme of the present invention.
The preparation that embodiment 1 produces with seed culture of viruses
1.1 a little less than the causing of canine distemper virus (CDV-YB) separated strain
Isolating canine distemper virus (CDV-YB strain) was passaged to for 70 generations through vaccinization CEF (CEF), changes afterwards at the Vero cell and uploaded for 20 generations, continue then to be uploaded to for 105 generations at CEF (CEF).In the process of going down to posterity, the titre of virus improves gradually, and strain more and more adapts to CEF and Vero cell.But virus lowers along with the increase of passage number the pathogenicity of dog gradually; Tests such as specificity through morphology evaluation, the check of pure property, virus and exogenous virus check, result verification the CDV-YB low virulent strain (CDV-YBR) cultivated have typical paramyxovirus particle characteristics (see figure 1), pure no exogenous virus pollutes; Can find out that from the immuning effect test result CDV-YBR strain can provide good protection to the dog of homology strong virus attack.
1.2 Canine Parvovirus CPV-YN) a little less than the causing of separated strain
Through continuous synchronization inoculation CRFK passage to 110 generation, in the process of going down to posterity, the titre of virus improves gradually with isolating Canine Parvovirus (CPV-YN strain), and strain more and more adapts to the CRFK cell.In contrast, virus lowers along with the increase of passage number the pathogenicity of dog gradually; Through morphology identify, result of the tests such as the specificity of pure property check, virus and exogenous virus check have verified that the CPV-YN low virulent strain of cultivating (CPV-YNR) has typical parvovirus particle characteristics (see figure 2), pure no exogenous virus pollutes; Can find out that from the immuning effect test result CPV-YNR strain can provide good protection to the dog of homology strong virus attack.
1.3 a little less than the causing of hepatitis infectiosa canis virus I type (CAV-H) separated strain
Isolating hepatitis infectiosa canis virus (CAV-H strain) was passaged to for 120 generations through the vaccinization mdck cell, and in the process of going down to posterity, the titre of virus improves gradually, and strain more and more adapts to mdck cell.In contrast, virus lowers along with the increase of passage number the pathogenicity of dog gradually; Through morphology identify, result of the tests such as the specificity of pure property check, virus and exogenous virus check have verified that the CAV-H low virulent strain of cultivating (called after CAV-HR) has typical adenovirion characteristic (see figure 3), pure no exogenous virus pollutes; Can find out that from the immuning effect test result CAV-HR strain a little less than causing can provide good protection to the dog of homology strong virus attack.
Embodiment 2 seedlings are prepared with viral liquid
2.1CDV-YBR the preparation of cell venom
Select well-grown CEF monolayer, remove culture fluid after, change containing the cell maintenance medium of 2% seed culture of viruses adjustment pH value 7.2～7.4.Put 33 ℃ of cultivations.Every day, observation of cell found that the cell of growth failure or pollution should be discarded immediately.Results are put-20 ℃ of preservations when CPE reaches 70%.
2.2CPV-YNR the preparation of cell venom
When CRFK goes down to posterity, press 3% inoculation CPV-YNR seed culture of viruses of growth-promoting media, adjustment pH value 7.2～7.4 is put 37 ℃ of cultivations.Every day, observation of cell found that the cell of growth failure or pollution should be discarded immediately.Results are put-20 ℃ of preservations when CPE reaches 70%.
2.3CAV-HR the preparation of cell venom
Select well-grown MDCK monolayer, change to contain the cell maintenance medium of 2% seed culture of viruses.Adjustment pH value 7.2～7.4 is put 37 ℃ of cultivations.Every day, observation of cell found that the cell of growth failure or pollution should be discarded immediately.Results are put-20 ℃ of preservations when CPE reaches 70%.
The preparation of embodiment 3 canine distemper, parvovirus, adenovirus I type trigeminal live vaccine
The viral liquid of the different content that embodiment 2 is prepared and stabilizing agent after fully shaking up, quantitatively are sub-packed in the sterilization vaccine bottle after adding the freeze drying protectant lyophilizing by 9: 1 volume ratio; After packing finishes, move into immediately in the freeze dryer, put earlier below-36 ℃ pre-freeze 3-4 hour; The turn on pump distillation is 16-18 hour then, when temperature is raised to 26-27 ℃, keeps after 4-5 hour and closes pump, finds time; And under vacuum state automatic stopper adding, gland, accomplish lyophilizing.
Test Example 1 canine distemper virus, parvovirus and adenopathy viral disease trigeminal live vaccine immuning effect test
1 material and method
1.1 test is with vaccine canine distemper disease, parvovirus infection, adenopathy viral disease trigeminal live vaccine; Method for preparing according to embodiment 1-3 is prepared into 5 batches, and lot number is 9701,9702,9703,9704,9705.
1.2 2～4 monthly age of laboratory animal healthy dogs, its CDV, CPV, CAV serum neutralizing antibody all≤1: 2.
1.3 attack with strong malicious CDV-YB (8 generation), CPV-YN (7 generation), CAV-H (8 generation).
1.4 every batch of vaccine of test method dilutes with water for injection, 2～4 monthly age of intramuscular inoculation 15 of dogs, 1 part/dog.15 dogs are divided into 3 groups after 21 days, and 5 every group, every group of dog attacked a kind of strong poison, and dosage is 200TCID
50/ dog.Establish contrast 5 of dogs (injecting normal saline 1ml) for every group simultaneously.Each organizes the equal isolated rearing of dog, observes 21 days, and morbidity and the death condition of dog respectively organized in record.
2 results
2.1CDV counteracting toxic substances protection result with 5 batches of vaccines to 2～4 the monthly age dog carry out immunity, back 21 days of immunity is with 200TCID
50The CDV-YB of/dog dosage is strong, and poison is attacked, and the protective rate of 9701 batches of vaccine immunity dogs is 4/5, and the protective rate of other batches is 5/5, and matched group 5/5 morbidity has 2 death after 4 days in the morbidity dog.The result sees table 1.
Table 1 CDV counteracting toxic substances protection result
2.2CPV counteracting toxic substances protection result to 5 batches of vaccines to 2～4 the monthly age dog carry out immunity, 1 part/dog, back 21 days of immunity is with 200TCID
50The CPV-YN of/dog dosage is strong, and poison is attacked, and the protective rate of 9702,9704 batches of vaccine immunity dogs is 4/5, and other batches are 5/5, and matched group 5/5 morbidity has 1 dog death, and the result sees table 2.
Table 2 CPV counteracting toxic substances protection result
2.3CAV counteracting toxic substances protection result to 5 batches of vaccines to 2～4 the monthly age dog carry out immunity, 1 part/dog, back 21 days of immunity is with 200TCID
50The CAV-H of/dog dosage is strong, and poison is attacked, and the protective rate of 9701 batches of vaccine immunity dogs is 4/5, and other batches are 5/5, and matched group 5/5 morbidity has 2 dogs dead at 4-5 days cameras in the morbidity dog.The result sees table 3.
Table 3 CAV counteracting toxic substances protection result
3 conclusion (of pressure testing)s canine distemper of the present invention, parvovirus infection, adenopathy viral disease trigeminal live vaccine all reach 4/5 to the protective rate of canine distemper, parvovirus infection, adenopathy viral disease, have good protection effect.
Claims (7)
1. a canine distemper (Canine distemper virus), Canine Parvovirus (Canine parvovirus) and I type hepatitis infectiosa canis virus (Canine adenovirus) three attenuated live vaccines; Comprise: microbial preservation number is the canine distemper attenuated vaccine strain of CGMCC No.3810, and microbial preservation number is that Canine Parvovirus attenuated vaccine strain and the microbial preservation of CGMCC No.3841 number is the I type canine adenovirus attenuated vaccine strain of CGMCC No.3809.
2. according to described three attenuated live vaccines of claim 1, it is characterized in that: said canine distemper attenuated vaccine strain, the malicious valency of Canine Parvovirus attenuated vaccine strain or I type canine adenovirus attenuated vaccine strain is more than or equal to 10
4.5TCID
50/ head part.
3. the method for preparing of the said canine distemper of claim 1, Canine Parvovirus and hepatitis infectiosa canis virus I type virus three attenuated live vaccines; Comprise: (1) cultivating microorganism preserving number respectively is the canine distemper attenuated vaccine strain of CGMCC No.3810; Microbial preservation number is that Canine Parvovirus attenuated vaccine strain and the microbial preservation of CGMCC No.3841 number is the I type canine adenovirus attenuated vaccine strain of CGMCC No.3809, obtains the viral liquid of above-described each attenuated vaccine strain; (2) respectively the viral liquid of each attenuated vaccine strain is carried out titration of virus, the viral liquid of each attenuated vaccine strain is mixed, filter, add freeze drying protectant, lyophilizing promptly gets;
The compound method of wherein said freeze drying protectant comprises: (1) gelatin: 8g; Sucrose: 40g; Deionized water: 100ml; The adjustment pH value is 6.8～7.0; (2) 116 ℃ of sterilization 40min promptly get.
4. according to the method for preparing of the said canine distemper of claim 3, Canine Parvovirus and hepatitis infectiosa canis virus I type virus three attenuated live vaccines; It is characterized in that the cultivation of said canine distemper attenuated vaccine strain comprises: select through aseptic well-grown CEF monolayer that detection of mycoplasma is negative; After removing culture fluid; Change to contain the cell maintenance medium of 2% seed culture of viruses, adjustment pH value 7.2～7.4 is put 33 ℃ of cultivations; Results when CPE reaches 70%.
5. according to the method for preparing of the said canine distemper of claim 3, Canine Parvovirus and hepatitis infectiosa canis virus I type virus three attenuated live vaccines; It is characterized in that; The cultivation of said Canine Parvovirus attenuated vaccine strain comprises: seed culture of viruses is inoculated in through aseptic by 3% of cell maintenance medium amount synchronously; Well-grown CRFK monolayer that detection of mycoplasma is negative, the adjustment pH value is 7.2～7.4, puts 37 ℃ of cultivations; Treat that 70% cell CPE occurs and can gather in the crops.
6. according to the method for preparing of the said canine distemper of claim 3, Canine Parvovirus and hepatitis infectiosa canis virus I type virus three attenuated live vaccines; It is characterized in that: the cultivation of hepatitis infectiosa canis virus I type attenuated vaccine strain comprises: select through aseptic; Well-grown MDCK monolayer that detection of mycoplasma is negative; After removing culture fluid, change to contain the cell maintenance medium of 2% seed culture of viruses, adjustment pH value 7.2～7.4; Put 37 ℃ of cultivations; Results when CPE reaches 70%.
7. according to the method for preparing of the said canine distemper of claim 3, Canine Parvovirus and hepatitis infectiosa canis virus I type virus three attenuated live vaccines, it is characterized in that: mixed viral liquid is mixed with the volume ratio of freeze drying protectant by 9:1.
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| CN102807970B (en) * | 2012-05-17 | 2013-12-25 | 中国农业科学院哈尔滨兽医研究所 | Canine distemper virus (CDV) sensitive cell line and establishment method and application thereof |
| CN103937752B (en) * | 2014-03-03 | 2016-08-31 | 中国农业科学院哈尔滨兽医研究所 | A kind of H3N2 hypotype canine influenza virus inactivated vaccine and its preparation method and application |
| CN107320720A (en) * | 2016-04-29 | 2017-11-07 | 普莱柯生物工程股份有限公司 | A kind of vaccine combination, kit and application |
| CN108066758B (en) * | 2017-12-26 | 2020-01-10 | 中国农业科学院特产研究所 | Raccoon dog canine distemper-parvovirus enteritis bigeminal live vaccine and preparation method and application thereof |
| CN110841064B (en) * | 2019-11-07 | 2020-11-27 | 衡阳师范学院 | Canine adenovirus type I inactivated vaccine and preparation method thereof |
| CN112316128B (en) * | 2020-10-20 | 2021-08-24 | 辽宁益康生物股份有限公司 | Canine distemper virus and canine parvovirus infectious hepatitis virus triple live vaccine |
| CN112370531B (en) * | 2020-11-18 | 2022-09-23 | 金宇保灵生物药品有限公司 | Heat-resistant protective agent for canine distemper, canine parvovirus, canine adenovirus and canine parainfluenza four-combined live vaccine, and preparation method and application thereof |
| CN112725288B (en) * | 2021-01-15 | 2022-07-26 | 北京华夏兴洋生物科技有限公司 | Canine adenovirus type 2 attenuated vaccine strain and application thereof |
| CN117106732B (en) * | 2023-09-07 | 2024-02-06 | 华中农业大学 | Canine parvovirus and application thereof |
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Inventor after: Liu Dafei Inventor after: Jiang Yitong Inventor after: Zhang Hongying Inventor after: Gao Yan Inventor after: Liu Dacheng Inventor before: Zhang Hongying Inventor before: Liu Dafei Inventor before: Qu Liandong Inventor before: Wang Mouping Inventor before: Liu Likui |
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