CN1137347A - Canine penton (canine yabies, pestilence, parvovirus, adenovirus No.2 type and parainfluenza) live vaccine - Google Patents
Canine penton (canine yabies, pestilence, parvovirus, adenovirus No.2 type and parainfluenza) live vaccine Download PDFInfo
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- CN1137347A CN1137347A CN 95106744 CN95106744A CN1137347A CN 1137347 A CN1137347 A CN 1137347A CN 95106744 CN95106744 CN 95106744 CN 95106744 A CN95106744 A CN 95106744A CN 1137347 A CN1137347 A CN 1137347A
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Abstract
The canine penton (canine rabies, pestilence, parvovirus, No.2 type adenovirus and parainfluenza) live vaccine is made up by using wild virus obtained from Chinese diseased dog body and domesticated canine distemper attenuated virus strain as object, and using canine rabies virus, canine parvovirus, canine No.2 type adenovirus and canine parainfluenza virus which are imported from abroad, adding a proper quantity of heat-resistant stabilizing agent and adopting such new techniques as temp.-converting multiplication and physical freeze concentration. The invented canine penton live vaccine is high in vaccine concentration, good in immunological effect, simple in production process and low in cost.
Description
The present invention relates to adopt that canine parainfluenza virus, CDV, rabies virus, parvovirus, adenovirus make is the pharmaceutical products of viral antigen.
Rabies, canine distemper, canine parvovirus enteritis, HCC/canine infectious laryngotracheitis and dog parainfluenza are six common among the dog group big infectious diseases, the eqpidemic disease incidence and mortality that wherein has has had a strong impact on development and guarantor's kind of famous and precious dog kind and the life security of the antitheft dog of keeping goal among the people of army dog police dog up to 50-100%.States such as external American and Britain, method, day, Russia have just begun to develop the individual event live vaccine of the above six kinds of dog infectious diseases of prevention in the seventies, to the later stage eighties, on the basis of tetrad (not having mad dog) seedling, succeeded in developing the canine penton seedling that comprises the Leptospira rhzomorph.Though there is veterinary rabies sheep brain inactivated vaccine in the mid-1960s China, because of complex process, dosage is big, side effect is strong, cost is high, be difficult to satisfy epidemic prevention needs in all parts of the country.Nineteen eighty-three, academy of agricultural sciences China Veterinary Drugs Supervisory Inst. introduced mad dog ERA attenuated strain from external, and had trial-produceed the veterinary rabies live vaccine successfully with relevant producer cooperation.Especially China is still passive in the seed poison of China's native country seedling so far for canine distemper, and still has skew with the external wild malicious antigenicity of vaccine and China of planting the poison preparation, and the result has caused partly using the protection of animals of seedling unit uncertain.
The objective of the invention is to adopt wild poison and the canine distemper attenuated strain through causing weak domestication available from getting in the sick dog body of China is object, and and be mixed with the 5-linked seedling together from rabies virus, canine parvovirus, hepatitis infectiosa canis virus 2 types, the canine parainfluenza virus of external introduction, and add suitable heat-resisting stabilizing agent and make.This 5-linked seedling safety is reliable, good immune effect and with strong points, is fit to the prevention of the sick dog disease of China, can be used for preventing rabies, canine distemper, canine parvovirus, HCC/canine infectious laryngotracheitis and six kinds of infectious diseases of dog parainfluenza; And the seedling craft science, program is workable, and economic benefit is considerable, is suitable for suitability for industrialized production.
The object of the present invention is achieved like this: canine penton (the mad dog of dog, canine distemper, canine parvovirus, hepatitis infectiosa canis virus 2 types, dog parainfluenza) live vaccine.The main flow process of its production technology is: plant the malicious valency of the domestication of going down to posterity on malicious isolation identification → cell → temperature-speed-changing amplification → receipts poison → safety, efficacy test → concentrate → measure → join in proportion seedling → add stabilizing agent → packing → freeze-drying → product inspection → labeling packing.Be the seed culture of viruses of the existing seedling of domestic majority with kind of the rabies poison strain ERA-836 of poison wherein as seedling, canine parvovirus CPV-XN
1, hepatitis infectiosa canis virus 2 type CAV
2-XN
3With canine parainfluenza virus CPIV-XN
4Three strain virus are by isolation identification screening the dog tetrad seedling of producing from the SoLovary company of external introduction and get, its special feature is: a strain wild poison of the CDV attenuated strain among the present invention for getting in the sick dog body in NORTHWEST CHINA area, get through the domestication of going down to posterity on the different cell, clonal expansion, mutagenesis attenuation, screening being arranged in provenance, through identifying, this strain domestication spectrum clear (seeing Table) genetic stability is reliable, have common antigen with international standard strain SH-CDVOnder-stepoot strain.Therefore, the optimal candidate strain of the weak poison strain of this CDV system preparation live vaccine.
In the production of vaccine process, the invention provides a kind of temperature-speed-changing rotating and culturing new technology.Inquiring into through theory may be in the virus of 35 ℃ of cultivations, along with propagation constantly is released in the culture fluid, this virus is because of fragile to photopigment and salt ion resistance, make virus constantly deactivation in liquid, so just reduced the malicious valency in the unit volume, reach at 30% o'clock if treat pathology, cool the temperature to 22 ℃ and increase rotating speed, treat to receive poison when pathology forms 80-90%, can improve malicious valency about 33% like this, average T CID
50Can improve 3 times.
The present invention also provides the freeze thawing of employing culture physical property to concentrate new technology.To tame the viral seedling liquid semi-finished product in back freezing after, the venom that will freeze moves under the room temperature again, makes it melt naturally, takes out district's carrying liqs of abandoning upper strata about 1/3 or 1/2, will remain venom and filter, gained filtrate is the concentrated former seedling of property of seedling usefulness.
The present invention also provides a kind of new heat-resisting stabilizing agent prescription, is used to protect the antigenicity of vaccine, sees Table II.
Accompanying drawing is the process flow diagram of canine penton vaccine.
Embodiment is described in further detail below in conjunction with (accompanying drawing).
From the dog tetrad seedling that the Solovary company of external introduction makes, separate the good CPV-XN of proliferative
1, CAV
2-XN
3, and CPIV-XN
4Strain also intersects by the heterogenous cell that exsomatizes in addition and goes down to posterity, and obtains the unique CPV-112 strain of China, and rabies virus ERA-836 then is a domestic current seedling attenuated strain.Be used for cultured chick embryo fibroblast (CEF) and prepare BHK voluntarily with SPF chicken embryo
21The strain hamster kidney cell, African green monkey kidney cell line Vero, F
81Strain cat kidney cell line and MDCK strain MDCK system provide by prison institute or Ministry of Public Health drug inspection office in the Ministry of Agriculture.With ERA-836, CPV-XN
1, CPV-XN
112, CAV
2-XN
2And CPIV-XN
4Respectively at BHK
21, F
81Cultivate on CFF, MDCK and the Vero cell.
Its process is respectively:
1. CDV-XN
112: connect on 5 generations of blind passage on the chick chorioallantoic membrane → CEF pass 47 generation → three time plaque colony screening kind poison → mdck cell and upload for 40 generations → CEF uploaded for 20 generations.When reaching for 90 generations, this strain promptly loses pathogenic to dog.
2. CPV-XN
1: at F
81Upload 1-5 generation.Be seedling canine parvovirus enteritis vaccine kind poison,
3. CAV
2-XN
3: upload 2-5 generation at MDCK, be dog hepatitis and laryngotracheitis vaccine seed poison.
4. CPIV-XN
4: increase foster 7 days at Vero and be dog secondary flow trouble seed poison.
5. ERA-836 is provided by the biological pharmaceutical factory of Lanzhou animal doctor early stage, and the later stage is at BHK
21Cultivate preparation on the cell and get.
In incubation,, just blake bottle rev/min is transferred under 22 ℃, 14 rev/mins of conditions of rotating speed, with the malicious valency in the raising unit volume by 35 ℃, rotating speed 8-9 as the CPE of virus when reaching 30%.
After finishing, incubation measures safety, effectiveness and the viral antigen titre of vaccine, the blake bottle of singly planting vaccine will be housed then to be descended freezing 12 hours at-30 ℃, then blake bottle is moved under the room temperature, after treating that bottle interior venom melts naturally, take out and abandon the about 1/2-1/3 liquid in upper strata, the venom of lower floor is the vaccine liquid after concentrating, after conversion is tired, and can be with ERA-636, CPV-XN
1, CDV-XN
112, CAV
2-XN
3And CPIV-XN
4Culture fluid in 0.15,0.40,0.15 and the ratio of 0.15ml mix, with malicious seedling liquid and heat-resisting stabilizing agent mixed in equal amounts a little less than this, fully shake up, after copper mesh filters, quantitatively be sub-packed in the bottle, every bottle of 2ml puts lyophilization in the freeze dryer of tamponade automatically, measure its vacuum water content at last, and do three inspection backs in low temperature or 4-8 ℃ of preservation.
Vaccine is made aseptic, safety and efficacy determinations.(seeing Table III).
Advantage of the present invention:
1, with strong points:
Produce canine distemper attenuated strain used in the canine penton seedling in the sick dog body of China, getting, be suitable for six kinds of infectious diseases of the sick dog of China with four strain strains of import by the proper proportion proportioning: rabies, canine distemper, canine parvovirus enteritis, HCC/canine infectious laryngotracheitis, the para-influenzal prevention of dog are strictly the first-selected special effect medicine of the sick dog prevention of infectious disease of China.
2, protectiveness is good:
Heat-resistant stability prescription instant capacity used in this canine penton seedling is good, and warm tolerance is strong, and outward appearance can be compared U.S. with the import vaccine.
3, malicious valency is renderd a service high:
Adopt to change warm rotating speed in the production technology and cultivate and two kinds of new technologies of freeze concentration, make that average malicious valency improves about 1～2 times in the vaccine, and keep the stability of Proantigen titre, can keep the integrality of viral disease virion a little less than in the of five kinds.What deserves to be mentioned is that this technology also can reject the calf serum in the seedling liquid, the foreign protei allergy of avoiding dog body inoculation back to be produced takes place, and unit volume inner virus content is improved exponentially.
Production technology science of the present invention, equipment is less demanding, can be suitable for industrialized mass, thereby this canine penton height of seedling tires honest and cleanly, helps generally applying in China.
Table I CDV-XN
112Strain strain domestication spectrum
The propagation suckling mouse is refreshing to return the ※ generation propagation substrate temperature CPE TCID that goes down to posterity
50Through 35 ℃ of non-purulence ※ 1 of virulence Electronic Speculum animal 1 generation chicken embryo suede
-1Two-phase heat
35 ℃ 10 of hair chorioallantoic membrane plaque ± 2 generation chicken embryo suede
-2± tired
35 ℃ 10 of hair chorioallantoic membrane 3 generation chicken embryo suede
-3+ tired
Hair chorioallantoic membrane 4-5 becomes 33 ℃+10 for the chicken embryo
-3.510
-4+ tired
Fibrocyte 6-10 is for plaque clone 33 ℃+10
-410
-3+ one crosses type
(Vero) 11-30 is for stigma amplification 33 ℃+10
-410
-3.5+ subtract and ate one day
(Vero) 31-50 is for stigma amplification 33 ℃+10
-4.510
-3+ one crosses type
(Vero) 51-80 is for dog MDCK 33 ℃+10
-5.010
-4+ no abnormal
Cell 81-100 is for dog MDCK 33 ℃+10
-5.510
-3+ no abnormal
Cell 101-110 becomes 33 ℃+10 for the chicken embryo
-4.510
-2+ no abnormal
Fibrocyte
The heat-resisting protective agent prescription that Table II canine penton live vaccine is used
| Composition | Skim milk powder | Three heat up in a steamer water | Sucrose | Gelatin | ? ??VC | Sodium glutamate | Amikacin | Anphotericin | Powerful detoxifcation is quick | ? ??K2HO4 | ? ??KH2PO4 |
| Content | 800～ 1200(g) | 600～ 9000(ml) | 3％～ 10％ | 0.1～ 0.4％ | 1∶100～ 1∶200 | ?6％～ 10％ | 100～200 (ten thousand u) | 3.25～ 6.25(mg) | 50～200 (milliliter/ten thousand) | 10～ 13.15(g) | 2.0～ 4.48(g) |
Table III 5-linked freeze-dried products assay
| Lot number | Physical behavior | The bacterium inspection | Safety check | Efficacy test (17 days SN) | ||||||||
| Outward appearance | Dissolubility | Residual water (%) | Take off wall (%) | Vacuum | ??CPV | ?CDV | ?CAV 2 | CPIV | ??ERA | |||
| ?9401 ?9402 | The spongy dry agglomerate of milky | Dissolving rapidly | ≤3 | ??80 | Qualified | Aseptic | 4/4 (one) | ??＞1024 | ??＞128 | ??＞128 | ??＞128 | ??＞32 |
| ≤3 | ??90 | Qualified | ??＞512 | ??＞256 | ??＞128 | ??＞256 | ??＞64 | |||||
| ?9403 | ≤3 | ??90 | Qualified | ??# | ???#?# | ??＞512 | ??＞64 | ??＞256 | ??＞64 | ??＞32 | ||
| ?9404 | ≤3 | ??90 | Qualified | ??# | ???#?# | ??＞1024 | ??＞128 | ??＞256 | ??＞128 | ??＞128 | ||
| ?9405 | ≤3 | ??95 | Qualified | ??# | ???#?# | ??＞1024 | ??＞256 | ??＞256 | ??＞128 | ??＞64 | ||
| ?9406 | ≤3 | ??95 | Qualified | ??# | ???#?# | ??＞2048 | ??＞512 | ??＞128 | ??＞512 | ??＞32 | ||
| ?9409 | ≤3 | ??85 | Qualified | ??# | ???#?# | ??＞1024 | ??＞256 | ??＞256 | ??＞256 | ??＞32 | ||
| ?9410 | ≤3 | ??85 | Qualified | ??# | ???#?# | ??＞512 | ??＞128 | ??＞128 | ??＞256 | ??＞64 | ||
| ?9411 | ≤3 | ??85 | Qualified | ??# | ???#?# | ??＞512 | ??＞256 | ??＞256 | ??＞256 | ??＞32 | ||
| ?9412 | ≤3 | ??100 | Qualified | ??# | ???#?# | ??＞1024 | ??＞512 | ??＞512 | ??＞512 | ??＞32 | ||
| ?9416 | ≤3 | ??100 | Qualified | ??# | ???#?# | ??＞1024 | ??＞512 | ??＞256 | ??＞512 | ??＞32 | ||
| ?9418 | ≤3.5 | ??90 | Qualified | ??# | ???#?# | ??＞512 | ??＞256 | ??＞128 | ??＞256 | ??＞64 | ||
Claims (4)
1. canine penton (the mad dog of dog, canine distemper, canine parvovirus, hepatitis infectiosa canis virus 2 types, dog parainfluenza) live vaccine, the main flow process of its production technology is: plant the malicious valency of the domestication of going down to posterity on malicious isolation identification → cell → temperature-speed-changing amplification → receipts poison → safety, efficacy test → concentrate → measure → join in proportion seedling → add stabilizing agent → packing → freeze-drying → product inspection → labeling packing.Be the seed culture of viruses of the existing seedling of domestic majority with kind of the rabies poison strain ERA-836 of poison wherein as seedling, canine parvovirus CPV-XN
1, hepatitis infectiosa canis virus 2 type CAV
2-XN
3With canine parainfluenza virus CPIV-XN
4The all isolation identification screenings and getting from the dog tetrad seedling that the SoLovary company of external introduction produces of three strain virus, it is characterized in that a strain wild poison of the sick and weak strain of canine distemper among the present invention in the sick dog body in NORTHWEST CHINA area, getting, get through the domestication of going down to posterity on the different cell, clonal expansion, mutagenesis attenuation, screening being arranged in provenance, through identifying, this strain domestication spectrum is clear, genetic stability is reliable, have common antigen with international standard strain SH-CDV strain, the optimal candidate strain of the weak poison strain of this CDV system preparation live vaccine.
2. canine penton seedling according to claim 1, it is characterized in that: in the production of vaccine process, the invention provides a kind of temperature-speed-changing and cultivate new technology, treat in incubation that promptly cytopathy reaches at 30% o'clock, temperature downgraded and increase rotating speed, treat to receive when pathology forms 80-90% poison, can improve malicious valency about 33% like this, average T CID
50Can improve 3 times.
3. canine penton seedling according to claim 1 and 2, it is characterized in that: the present invention also provides the freeze thawing of a kind of culture physical property to concentrate new technology in the production of vaccine process, with viral seedling liquid semi-finished product freezing after, the venom that will freeze moves under the room temperature again, make it melt naturally, take out district's carrying liqs of abandoning upper strata about 1/3 or 1/2, will remain venom and filter, gained filtrate is seedling with concentrating the former seedling of property.
4. canine penton seedling according to claim 3 is characterized in that: the present invention also provides a kind of new heat-resisting stabilizing agent prescription, is used to protect the antigenicity of vaccine,
Its prescription is: skim milk powder 800～1200g, three heat up in a steamer water 600～9000ml, sucrose 3%～10%, gelatin 0.1～0.4%, VC 1: 100～1: 200, sodium glutamate 6%～10%, amikacin 100～2,000,000 μ, anphotericin 3.25～6.25mg, powerful detoxifcation is quick 50～200 milliliters/ten thousand, K
2HO
410～13.15g, KH
2PO
42.0～4.48g.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 95106744 CN1137347A (en) | 1995-06-22 | 1995-06-22 | Canine penton (canine yabies, pestilence, parvovirus, adenovirus No.2 type and parainfluenza) live vaccine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 95106744 CN1137347A (en) | 1995-06-22 | 1995-06-22 | Canine penton (canine yabies, pestilence, parvovirus, adenovirus No.2 type and parainfluenza) live vaccine |
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| Publication Number | Publication Date |
|---|---|
| CN1137347A true CN1137347A (en) | 1996-12-11 |
Family
ID=5076009
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 95106744 Pending CN1137347A (en) | 1995-06-22 | 1995-06-22 | Canine penton (canine yabies, pestilence, parvovirus, adenovirus No.2 type and parainfluenza) live vaccine |
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|---|---|
| CN (1) | CN1137347A (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7682619B2 (en) | 2006-04-06 | 2010-03-23 | Cornell Research Foundation, Inc. | Canine influenza virus |
| CN102488894A (en) * | 2011-12-31 | 2012-06-13 | 齐鲁动物保健品有限公司 | Fox encephalitis live vaccine and preparation method thereof |
| CN103740862A (en) * | 2013-12-30 | 2014-04-23 | 中华人民共和国沈阳出入境检验检疫局 | Multi-PCR (Polymerase Chain Reaction) detecting primer group for canine distemper virus, canine adenovirus type 2 and canine parainfluenza virus |
| CN101573137B (en) * | 2006-12-27 | 2014-05-14 | 硕腾P有限责任公司 | Methods of vaccine administration |
| CN105884889A (en) * | 2016-04-18 | 2016-08-24 | 内蒙古神元生物工程股份有限公司 | Canine distemper/infectious hepatitis/parvovirus triple egg yolk antibody and preparation method thereof |
| CN107537033A (en) * | 2016-06-23 | 2018-01-05 | 普莱柯生物工程股份有限公司 | Vaccine combination, kit and its application |
| CN107988174A (en) * | 2017-12-13 | 2018-05-04 | 新疆农业大学 | A kind of rabies-canine distemper-the strain of canine parvovirus gene recombined virus, construction method and its application |
| CN110302374A (en) * | 2019-08-09 | 2019-10-08 | 中国农业科学院特产研究所 | Canine vaccine and its preparation method and application |
| CN112316128A (en) * | 2020-10-20 | 2021-02-05 | 辽宁益康生物股份有限公司 | Canine distemper virus and canine parvovirus infectious hepatitis virus triple live vaccine |
-
1995
- 1995-06-22 CN CN 95106744 patent/CN1137347A/en active Pending
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7682619B2 (en) | 2006-04-06 | 2010-03-23 | Cornell Research Foundation, Inc. | Canine influenza virus |
| CN101573137B (en) * | 2006-12-27 | 2014-05-14 | 硕腾P有限责任公司 | Methods of vaccine administration |
| CN102488894A (en) * | 2011-12-31 | 2012-06-13 | 齐鲁动物保健品有限公司 | Fox encephalitis live vaccine and preparation method thereof |
| CN103740862A (en) * | 2013-12-30 | 2014-04-23 | 中华人民共和国沈阳出入境检验检疫局 | Multi-PCR (Polymerase Chain Reaction) detecting primer group for canine distemper virus, canine adenovirus type 2 and canine parainfluenza virus |
| CN103740862B (en) * | 2013-12-30 | 2015-10-28 | 中华人民共和国沈阳出入境检验检疫局 | The multiple PCR detection primer group of canine distemper virus, hepatitis infectiosa canis virus II type and canine parainfluenza virus |
| CN105884889A (en) * | 2016-04-18 | 2016-08-24 | 内蒙古神元生物工程股份有限公司 | Canine distemper/infectious hepatitis/parvovirus triple egg yolk antibody and preparation method thereof |
| CN107537033A (en) * | 2016-06-23 | 2018-01-05 | 普莱柯生物工程股份有限公司 | Vaccine combination, kit and its application |
| CN107988174A (en) * | 2017-12-13 | 2018-05-04 | 新疆农业大学 | A kind of rabies-canine distemper-the strain of canine parvovirus gene recombined virus, construction method and its application |
| CN110302374A (en) * | 2019-08-09 | 2019-10-08 | 中国农业科学院特产研究所 | Canine vaccine and its preparation method and application |
| CN110302374B (en) * | 2019-08-09 | 2023-04-14 | 黑龙江八一农垦大学 | Canine vaccine and preparation method and application thereof |
| CN112316128A (en) * | 2020-10-20 | 2021-02-05 | 辽宁益康生物股份有限公司 | Canine distemper virus and canine parvovirus infectious hepatitis virus triple live vaccine |
| 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 |
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