CN1546658A - Artificially recombinant H9N2 hypotype influenza virus and its uses - Google Patents
Artificially recombinant H9N2 hypotype influenza virus and its uses Download PDFInfo
- Publication number
- CN1546658A CN1546658A CNA2003101077322A CN200310107732A CN1546658A CN 1546658 A CN1546658 A CN 1546658A CN A2003101077322 A CNA2003101077322 A CN A2003101077322A CN 200310107732 A CN200310107732 A CN 200310107732A CN 1546658 A CN1546658 A CN 1546658A
- Authority
- CN
- China
- Prior art keywords
- influenza virus
- subtype
- virus
- chicken
- vaccine
- 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.)
- Granted
Links
- 241000712461 unidentified influenza virus Species 0.000 title claims description 37
- 241001473386 H9N2 subtype Species 0.000 title claims description 29
- 241000287828 Gallus gallus Species 0.000 claims abstract description 25
- 230000006798 recombination Effects 0.000 claims abstract description 17
- 238000005215 recombination Methods 0.000 claims abstract description 16
- 241001500351 Influenzavirus A Species 0.000 claims abstract description 10
- 210000001161 mammalian embryo Anatomy 0.000 claims description 16
- 229960005486 vaccine Drugs 0.000 claims description 14
- 206010022000 influenza Diseases 0.000 claims description 7
- 241001465754 Metazoa Species 0.000 claims description 6
- 101150080862 NA gene Proteins 0.000 claims description 5
- 230000007918 pathogenicity Effects 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 abstract description 8
- 241000700605 Viruses Species 0.000 description 21
- 101710154606 Hemagglutinin Proteins 0.000 description 9
- 101710093908 Outer capsid protein VP4 Proteins 0.000 description 9
- 101710135467 Outer capsid protein sigma-1 Proteins 0.000 description 9
- 101710176177 Protein A56 Proteins 0.000 description 9
- 239000000185 hemagglutinin Substances 0.000 description 9
- 108010006232 Neuraminidase Proteins 0.000 description 5
- 102000005348 Neuraminidase Human genes 0.000 description 5
- 230000003612 virological effect Effects 0.000 description 4
- 210000003837 chick embryo Anatomy 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 208000002979 Influenza in Birds Diseases 0.000 description 2
- 206010064097 avian influenza Diseases 0.000 description 2
- 238000009361 aviculture Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 230000035931 haemagglutination Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 230000008521 reorganization Effects 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 208000003322 Coinfection Diseases 0.000 description 1
- 241001500350 Influenzavirus B Species 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 241000712464 Orthomyxoviridae Species 0.000 description 1
- 241000710914 Totivirus Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 229940031551 inactivated vaccine Drugs 0.000 description 1
- 208000037797 influenza A Diseases 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 208000023504 respiratory system disease Diseases 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 238000012882 sequential analysis Methods 0.000 description 1
Abstract
The invention relates to an artificial recombination N9 hypotype influenza virus A/Harbin/Re-2/2003 (H9N2), artificial recombination H9 hypotype influenza virus A/Harbin/Re-2/2003(H9N2) characterized in that, it includes a HA of H9N2 hypotype fowl influenza virus A/chicken/ Guangxi/ 10/99(H9N2) and six internal genes of influenza virus A/PR/8/34(H1N1), PB2, PB1, PA, NP, M and NS. its storage number is CCTCC-V200311.
Description
Technical field:
The present invention relates to a kind of H9 subtype influenza virus of A/Harbin/Re-2/2003 (H9N2) of artificial recombination, and should the application of virus aspect the vaccine of control animal and human class H9N2 subtype influenza.
Background technology:
(avian influenza virus AIV) is the important cause of disease of bird and some mammiferous respiratory system diseases to avian influenza virus.AIV belongs on taxonomy: viral boundary (Vira)----orthomyxoviridae family (Orthomyxoviridae)----Influenza Virus (Influenza Virus A and B)----avian influenza virus (avian influenza virus).
According to surface protein antigenicity difference, avian influenza virus (avian influenza virus, AIV) be divided into 15 hemagglutinin (HA) and 9 neuraminidases (NA) hypotype, H9N2 hypotype AIV 1966 occurs in the turkey of the U.S. first first, nearly 10 years worldwide wild fowl and poultry in extensively distribute and popular.From pig, separated this subtype virus closely first in Hong Kong in 1998, in the body of 2 infants in Hong Kong, be separated to 2 strain H9N2 subtype influenza viruses, and indicated that this subtype avian influenza virus had huge public health meaning in 1999.Since nineteen ninety-six, China's aviculture is subjected to the puzzlement of H9 subtype avian influenza always.The totivirus inactivated vaccine of chicken embryo production makes the control of this disease obtain effect preferably in the application of aviculture.But, cause the vaccine of existing use to have shortcomings such as cost height, vaccine immunity phase weak point because the isolating street strain of terrain chicken embryo adaptability is relatively poor.
Summary of the invention:
The H9N2 subtype influenza virus that the purpose of this invention is to provide a kind of artificial recombination, according to the H9 subtype influenza virus of A/Harbin/Re-2/2003 (H9N2) of viral nomenclature rule called after artificial recombination, and should virus use aspect the vaccine of control animal and human class H9N2 subtype influenza.
Above-mentioned purpose realizes by following technical scheme:
The H9N2 subtype influenza virus of artificial recombination, have and the HA of H9N2 subtype avian influenza virus A/ chicken/Guangxi/10/99 (H9N2) and 6 internal gene PB2, PB1, PA, NP, M and the NS of NA gene and influenza virus A/PR/8/34 (H1N1), preserving number CCTCC-V200311.
The H9N2 subtype influenza virus of above-mentioned artificial recombination has cross-immunity widely to China's popular H9N2 avian influenza virus, has high titre chicken embryo adaptability, to chicken and chicken embryo had no pathogenicity.
The application of the H9N2 subtype influenza virus of above-mentioned artificial recombination aspect the vaccine of control animal and human class H9N2 subtype influenza.
This technical scheme has following beneficial effect:
1. influenza virus A/PR/8/34 (H1N1) is a laboratory chicken embryo adapted strain (PR8), promptly should virus can grow to very high titre in the chicken embryo, and this chicken embryo adaptability is by 6 internal gene decisions of virus.In addition, when cell of two influenza virus co-infected or chicken embryo, but the producer exchange, thus new gene recombined virus produced.Therefore, by the genetic recombination method, the HA of 6 internal gene of PR8 virus and epidemic strain, recombinating of NA gene can be obtained a new recombinant virus, this recombinant virus has and HA and the consistent antigenicity of NA genetic donor strain, also has the high titre growth characteristics of chicken embryo of PR8 virus simultaneously.
2. the H9N2 subtype influenza virus of the artificial reorganization of the present invention is HA and NA genetic donor with China avian influenza virus GX/99, and therefore, this virus is consistent with the most of H9N2 subtype avian influenza virus of the present popular of China antigenicity.Vaccine as vaccine strain is produced will produce good intersecting protective to most H9N2 subtype avian influenza virus epidemic strains.
3. the H9N2 subtype influenza virus of the artificial reorganization of the present invention contains 6 internal gene of the high titre adapted strain of chicken embryo PR8, the viral growth titre is compared with wild poison, can improve about 10 times, produces vaccine as vaccine strain, can reduce the production of vaccine cost greatly, improve vaccine quality.
The specific embodiment of the present invention:
Embodiment 1:
The H9N2 subtype influenza virus of artificial recombination is the H9 subtype influenza virus of A/Harbin/Re-2/2003 (H9N2) of artificial recombination, have and the HA of H9N2 subtype avian influenza virus A/ chicken/Guangxi/10/99 (H9N2) and 6 internal gene PB2, PB1, PA, NP, M and the NS of NA gene and influenza virus A/PR/8/34 (H1N1), preserving number CCTCC-V200311.
The H9N2 subtype influenza virus of above-mentioned artificial recombination has cross-immunity widely to China's popular H9N2 avian influenza virus, has high titre chicken embryo adaptability, to chicken and chicken embryo had no pathogenicity.
The application of a kind of H9N2 subtype influenza virus of above-mentioned artificial recombination aspect the vaccine of control animal and human class H9N2 subtype influenza.
The present invention is the made H9N2 hypotype of strain recombinant influenza A/ Harbin/Re-2/2003 (strain Re-2), and this virus HA and NA gene are from H9N2 subtype avian influenza virus A/ chicken/Guangxi/10/99 (GX/99), and 6 internal gene are from PR8 virus; This virus is consistent with China H9N2 subtype avian influenza virus epidemic strain antigenicity, and has good chicken embryo adaptability; Vaccine strain as anti-system H9 hypotype animal influenza.
The present invention is realized by following concrete steps:
Coinfection-two viral genome are recombinated: influenza virus A/PR/8/34 (PR8) (H1N1) with the domestic isolate A/chicken of H9N2 hypotype AIV/Guangxi/10/99 (GX/99) co-inoculation 10 age in days chick embryo allantoic cavities of crossing through UV treatment, hatch 24h for 35 ℃.
2. antibody screening-the remove virus of surperficial gene source: the allantoic fluid 50ul that under above-mentioned specific artificial condition, gathers in the crops in PR8 virus, mix with the serum of the anti-PR8 of 600ul of 4 doubling dilutions, hatch 30 minutes inoculated into chick embryo allantoic cavities afterwards under the room temperature, 35 ℃ hatch 48h after, chick embryo allantoic liquid carries out hemagglutination activity and detects, and the minimum and sample that have a hemagglutination activity of results serum dilution is used for second and takes turns screening.
3. dilution purifying-remove internal gene to derive from GX/99 virus: the virus that the same method screening obtains is carried out limited clone purification of 3 generations on the chicken embryo, utilizes fragments specific primer RT-PCR to identify and comes from GX/99,6 internal gene PB2, PB1, PA, NP, M and the NS recombinant virus from PR8 through sequential analysis screening HA and NA.
Claims (3)
1. the H9N2 subtype influenza virus of an artificial recombination, it is characterized in that: have and the HA of H9N2 subtype avian influenza virus A/ chicken/Guangxi/10/99 (H9N2) and 6 internal gene PB2, PB1, PA, NP, M and the NS of NA gene and influenza virus A/PR/8/34 (H1N1), preserving number CCTCC-V200311.
The H9N2 subtype influenza virus of 2 artificial recombination according to claim 1 is characterized in that: China's popular H9N2 avian influenza virus is had cross-immunity widely, have high titre chicken embryo adaptability, to chicken and chicken embryo had no pathogenicity.
The application of the H9N2 subtype influenza virus of 3 one kinds of claims 1 and 2 described artificial recombination aspect the vaccine of control animal and human class H9N2 subtype influenza.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200310107732 CN1263848C (en) | 2003-12-02 | 2003-12-02 | Artificially recombinant H9N2 hypotype influenza virus and its uses |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200310107732 CN1263848C (en) | 2003-12-02 | 2003-12-02 | Artificially recombinant H9N2 hypotype influenza virus and its uses |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1546658A true CN1546658A (en) | 2004-11-17 |
| CN1263848C CN1263848C (en) | 2006-07-12 |
Family
ID=34334459
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200310107732 Expired - Lifetime CN1263848C (en) | 2003-12-02 | 2003-12-02 | Artificially recombinant H9N2 hypotype influenza virus and its uses |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1263848C (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1326998C (en) * | 2004-11-25 | 2007-07-18 | 中国农业科学院哈尔滨兽医研究所 | Gene encoding hemagglutinin protein of H5 avian influenza virus and its application |
| CN1810961B (en) * | 2006-02-22 | 2010-05-12 | 中国人民解放军军事医学科学院微生物流行病研究所 | Recombinant influenza virus and its preparation and application |
| CN102399754A (en) * | 2011-05-27 | 2012-04-04 | 山东省农业科学院畜牧兽医研究所 | H9N2 avian influenza virus vaccine strain and application of H9N2 avian influenza virus vaccine strain in immune protection |
| CN104164408A (en) * | 2013-05-15 | 2014-11-26 | 普莱柯生物工程股份有限公司 | Newcastle disease, infectious bronchitis and avian influenza resisting vaccine composition and preparation |
| CN104232594A (en) * | 2014-09-11 | 2014-12-24 | 中国农业科学院哈尔滨兽医研究所 | Recombinant homologous avian H1N1 influenza virus inactivated vaccine strain (JS40/PR8) as well as preparation method and application of inactivated vaccine strain |
| CN104372023A (en) * | 2014-11-10 | 2015-02-25 | 中国农业大学 | Method for rescuing influenza virus |
| CN104611299A (en) * | 2014-09-25 | 2015-05-13 | 普莱柯生物工程股份有限公司 | Artificial recombinant H9N2 avian influenza virus strain, preparation method, vaccine composition and application thereof |
| CN104673759A (en) * | 2014-07-04 | 2015-06-03 | 中国农业科学院上海兽医研究所 | Exogenous gene-expressed recombinant influenza virus as well as preparation method and application of recombinant influenza virus |
| KR20220138290A (en) * | 2021-04-05 | 2022-10-12 | 대한민국(농림축산식품부 농림축산검역본부장) | H9N2 Strain Genetic Recombinant Low Pathogenic Avian Influenza A Virus, Manufacturing Method and Vaccine Composition Thereof Using H9N2 Avian Influenza Virus Belonging to Y280 Lineage |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101628932B (en) * | 2009-08-21 | 2012-06-06 | 中牧实业股份有限公司 | Bird flu synthetic peptide vaccine and preparation method thereof |
-
2003
- 2003-12-02 CN CN 200310107732 patent/CN1263848C/en not_active Expired - Lifetime
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1326998C (en) * | 2004-11-25 | 2007-07-18 | 中国农业科学院哈尔滨兽医研究所 | Gene encoding hemagglutinin protein of H5 avian influenza virus and its application |
| CN1810961B (en) * | 2006-02-22 | 2010-05-12 | 中国人民解放军军事医学科学院微生物流行病研究所 | Recombinant influenza virus and its preparation and application |
| CN102399754A (en) * | 2011-05-27 | 2012-04-04 | 山东省农业科学院畜牧兽医研究所 | H9N2 avian influenza virus vaccine strain and application of H9N2 avian influenza virus vaccine strain in immune protection |
| CN102399754B (en) * | 2011-05-27 | 2013-03-06 | 山东省农业科学院畜牧兽医研究所 | H9N2 avian influenza virus vaccine strain and application of H9N2 avian influenza virus vaccine strain in immune protection |
| CN104164408A (en) * | 2013-05-15 | 2014-11-26 | 普莱柯生物工程股份有限公司 | Newcastle disease, infectious bronchitis and avian influenza resisting vaccine composition and preparation |
| CN104673759A (en) * | 2014-07-04 | 2015-06-03 | 中国农业科学院上海兽医研究所 | Exogenous gene-expressed recombinant influenza virus as well as preparation method and application of recombinant influenza virus |
| CN104232594A (en) * | 2014-09-11 | 2014-12-24 | 中国农业科学院哈尔滨兽医研究所 | Recombinant homologous avian H1N1 influenza virus inactivated vaccine strain (JS40/PR8) as well as preparation method and application of inactivated vaccine strain |
| CN104611299B (en) * | 2014-09-25 | 2018-05-04 | 普莱柯生物工程股份有限公司 | H9N2 avian flus strain, preparation method, vaccine combination and its application of a kind of artificial recombination |
| CN104611299A (en) * | 2014-09-25 | 2015-05-13 | 普莱柯生物工程股份有限公司 | Artificial recombinant H9N2 avian influenza virus strain, preparation method, vaccine composition and application thereof |
| CN104372023A (en) * | 2014-11-10 | 2015-02-25 | 中国农业大学 | Method for rescuing influenza virus |
| CN104372023B (en) * | 2014-11-10 | 2017-02-08 | 中国农业大学 | Method for rescuing influenza virus |
| KR20220138290A (en) * | 2021-04-05 | 2022-10-12 | 대한민국(농림축산식품부 농림축산검역본부장) | H9N2 Strain Genetic Recombinant Low Pathogenic Avian Influenza A Virus, Manufacturing Method and Vaccine Composition Thereof Using H9N2 Avian Influenza Virus Belonging to Y280 Lineage |
| KR102556305B1 (en) | 2021-04-05 | 2023-07-19 | 대한민국 | H9N2 Strain Genetic Recombinant Low Pathogenic Avian Influenza A Virus, Manufacturing Method and Vaccine Composition Thereof Using H9N2 Avian Influenza Virus Belonging to Y280 Lineage |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1263848C (en) | 2006-07-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Choi et al. | Avian influenza viruses in Korean live poultry markets and their pathogenic potential | |
| Gambaryan et al. | Evolution of the receptor binding phenotype of influenza A (H5) viruses | |
| Liu et al. | The influenza virus gene pool in a poultry market in South central china | |
| Zhang et al. | Characterization of H9N2 influenza viruses isolated from vaccinated flocks in an integrated broiler chicken operation in eastern China during a 5 year period (1998–2002) | |
| CN1263848C (en) | Artificially recombinant H9N2 hypotype influenza virus and its uses | |
| Yassine et al. | Interspecies and intraspecies transmission of influenza A viruses: viral, host and environmental factors | |
| CN104232594B (en) | Recombination classes fowl type H1N1 inactivated influenza virus vaccines strain (JS40/PR8) and its preparation method and application | |
| CN1261564C (en) | Artificial recombined influenza virus and its application | |
| Zhu et al. | Comparison of biological characteristics of H9N2 avian influenza viruses isolated from different hosts | |
| CN103614345B (en) | A kind of influenza virus vaccine strain | |
| CN104844708A (en) | Horse anti-H7N9 subtype influenza specific immune globulin F(ab)<2> and preparation method thereo | |
| CN1746298B (en) | Artificial recombinant H7 subinfluenza virus and use thereof | |
| CN108300702B (en) | Chicken-derived H9N2 avian influenza virus cold-adapted strain screening method and application thereof | |
| CN102586195A (en) | Method for preparing avian influenza virus and inactivated vaccine thereof with Vero passage cells | |
| CN102223895A (en) | Method for production of ph stable enveloped viruses | |
| Marjuki et al. | Isolation of highly pathogenic avian influenza H5N1 virus from Saker falcons (Falco cherrug) in the Middle East | |
| CN102166276B (en) | Traditional Chinese medicine composition for preventing and treating influenza A and preparation method and application thereof | |
| RU2507256C2 (en) | Strain of influenza virus a/17/mallard/netherlands/00/95(h7n3) for production of live and inactivated influenza vaccines | |
| US4552757A (en) | Use in an animal host and precursors for vaccines utilizing avian-human reassortants to combat influenza A virus | |
| CN102839160A (en) | Recombination influenza virus of high-efficiency expression HA (Hyaluronic Acid) protein, as well as preparation method and application thereof | |
| CN103740654A (en) | Cold adaptation strain of influenza B virus Vero cell and application thereof | |
| TWI727638B (en) | Use of fucoidan for inhibiting avian influenza virus | |
| US20220118078A1 (en) | Production of viruses in continuously growing epithelial cell lines derived from chicken gut | |
| CN108103084A (en) | Distinguish immune and infection animal H5 subtype avian influenza vaccine strains and its preparation method and application | |
| CN102864127A (en) | Recombinant influenza virus for efficiently expressing HA (Hemagglutinin) protein and preparation method as well as 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 | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CX01 | Expiry of patent term | ||
| CX01 | Expiry of patent term |
Granted publication date: 20060712 |