CN102949720A - Biosecurity multifactor blood serum production method for African swine fever - Google Patents
Biosecurity multifactor blood serum production method for African swine fever Download PDFInfo
- Publication number
- CN102949720A CN102949720A CN2012103271218A CN201210327121A CN102949720A CN 102949720 A CN102949720 A CN 102949720A CN 2012103271218 A CN2012103271218 A CN 2012103271218A CN 201210327121 A CN201210327121 A CN 201210327121A CN 102949720 A CN102949720 A CN 102949720A
- Authority
- CN
- China
- Prior art keywords
- swine fever
- african swine
- serum
- manufacture method
- structural protein
- 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.)
- Pending
Links
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The invention relates to a biosecurity multifactor blood serum production method for African swine fever. The production method comprises the following steps: by adopting gene expression structure proteins P72, K205R, P54 and A104R, carrying out chemical purification, enveloping with incomplete Freund adjuvant, carrying out muscle immunity experiments on animal swine by three times, collecting swine blood after one month, separating the blood serum, carrying out serological test, subpackaging and storing.
Description
Technical field
The present invention relates to a kind of preparation method of serum, especially biological safety African swine fever antigen multiple-factor serum manufacture method.
Background technology
African swine fever is a kind of deadly infectious disease of the pig that caused by African swine fever virus, can cause swinery 100% M ﹠ M, is used for this disease of epidemic prevention without effective vaccine both at home and abroad at present, and this disease is classified as the category-A animal epidemic by International Office of Epizootics.This disease is mainly popular in Africa and European countries, and causes huge economic loss.In case morbidity can only be taked the measure of blocking, slaughtering and destroying.Last century the eighties, horse and African swine fever occurs for he finally causes its domestic then all live pigs all to be butchered burning.China has no precedent African swine fever, should maintain sharp vigilance for African swine fever, because the spread speed of African swine fever virus is fast, communication media is varied.Of particular concern is that in May, 2007, African swine fever has occured in Georgia, after this, in less than year, African swine fever has propagated into the former Soviet Union areas such as Armenia, Azerbaijan, Russia, has formed a new African swine fever epidemic regions in this area.Under present case, China should strengthen causing all pass in and out quarantine and sterilizations of African swine fever infection, and monitoring is flowed with Russian adjoining regional wild animal, carries out suitable African swine fever monitoring and Epidemiological study.
Diagnostic techniques is most important for the epidemic prevention of African swine fever, advanced diagnostic techniques helps to monitor for infected zone and surrounding area thereof, strict control African swine fever fast delimited infected area and vulnerable area from importing into overseas, carry out detailed Epidemiological study.External the most frequently used diagnostic techniques is ELISA, and the method both can detect serum, also can detect tissue fluid, can be used for diagnosis, monitoring and Epidemiological study.
Domestic existing African swine fever serology ELISA diagnostic techniques product without molding, main cause is for lacking positive control serum, because the serotype diagnostic kit of African swine fever at first needs positive serum is arranged as experiment contrast.On the one hand, African swine fever did not occur in China, the African swine fever virus that does not have laboratory to preserve; On the other hand, African swine fever is the category-A zoosis, at present also not from external introduction research virus.So domestic by experiment technology is produced artificial challenge's serum of African swine fever.Up to the present, the domestic African swine fever ELISA diagnostic kit that adopts all is that high price is from foreign procurement, the detection of every part of pig anteserum sample cost surpasses 200 yuans, the economic pressures that can bear considerably beyond this diagnostic reagent of use of raiser or even institute.
Biological safety African swine fever antigen multiple-factor serum thoroughly solves the bottleneck that China can't produce the African swine fever diagnostic kit, feasible method has two kinds at present, a kind of method is from giving a long price for positive serum abroad, another method is to prepare antigen by biotechnology, by animal inoculation, technology development have the height biological safety, cheaply, be suitable for the open positive control serum of diagnostic reagent.And first method, expensive except spending, to solve also that the bio-safety of bringing from external introduction African swine fever positive serum threatens and Trade technique barrier all unfavorable, be unsuitable for developing diagnostic reagent, and the latter be own innovative approach, design is unique, technology is rigorous, and is feasible in theory, and expressing protein has merged the existing basic condition of this laboratory and advantage, produce low-cost diagnosis and use positive control serum, then possess the genetic prerequisite of exploitation diagnostic kit.
Summary of the invention
One of purpose of the present invention is achieved through the following technical solutions:
Adopt four kinds of albumen of structural protein P72, K205R, P54, A104R of gene expression, through chemical purification, coated with freund 's incomplete adjuvant, time muscle immunization experiment animal pig gathers Sanguis sus domestica after one month, separation of serum in three batches, detect through serology, packing is preserved.
Structural protein P72, the K205R of four kinds of expression, P54, A104R Argine Monohydrochloride sequence are as follows:
1、P72
1 vsvegtsgpl lcnihdlhkp hqskpiltde ndtqrtcsht npkflsqhfpenshniqtag
61 kqditpitda tyldirrnvh yscngpqtpk yyqpplalwi klrfwfnenvnlaipsvsip
121 fgerfitikl asqkdlvnef pg
2、K205R
1 mvepreqffq dllsavdqqm dtvkndikdi mkektsfmvs fenfierydtmekniqdlqn
61 kyeemaanlm tvmtdtkiql gaiiaqleil mingtplpak kttikeamplpssntnneqt
121 sppasgktse tpkknptnam fftrsewass ntfrekfltp eiqaildeqfanktgierlh
181 aeglymwrtq fsdeqkkmvk emmkk
3、P54
1 mdseffqpvy prhygeclss tptpsffsth mytiliaivv liiiiivliy lfssrkkkaa
61 aaieeediqf inpyqdqqwa gvppqpgiak pagattgsvs kpvmdrpitn npvmdrpvtn
121 hpvtdrlvtd klgmatgepa aasapahsae pyttvttqnt asqtmsaienlrqrstythk
181 dlensl
4、A104R
1 mstkkkptit kqelyslvaa dtqlnkalie riftsqqkii qnalkhnqev iippgikftv
61 vtvkakparq ghnpatgepi qikakpehka vkiralkpvh dmln
The specific embodiment
Below the preferred embodiments of the present invention are described in detail; Should be appreciated that preferred embodiment only for the present invention is described, rather than in order to limit protection scope of the present invention.Adopt four kinds of albumen of structural protein P72, K205R, P54, A104R of gene expression, through chemical purification, coated with freund 's incomplete adjuvant, time muscle immunization experiment animal pig gathers Sanguis sus domestica after one month, separation of serum in three batches, detect through serology, packing is preserved.
<400> 1
1 vsvegtsgpl lcnihdlhkp hqskpiltde ndtqrtcsht npkflsqhfp enshniqtag
61 kqditpitda tyldirrnvh yscngpqtpk yyqpplalwi klrfwfnenv nlaipsvsip
121 fgerfitikl asqkdlvnef pg
<400> 2
1 mvepreqffq dllsavdqqm dtvkndikdi mkektsfmvs fenfierydt mekniqdlqn
61 kyeemaanlm tvmtdtkiql gaiiaqleil mingtplpak kttikeampl pssntnneqt
121 sppasgktse tpkknptnam fftrsewass ntfrekfltp eiqaildeqf anktgierlh
181 aeglymwrtq fsdeqkkmvk emmkk。
<400> 3
1 mdseffqpvy prhygeclss tptpsffsth mytiliaivv liiiiivliy lfssrkkkaa
61 aaieeediqf inpyqdqqwa gvppqpgiak pagattgsvs kpvmdrpitn npvmdrpvtn
121 hpvtdrlvtd klgmatgepa aasapahsae pyttvttqnt asqtmsaien lrqrstythk
181 dlensl
<400>4
1 mstkkkptit kqelyslvaa dtqlnkalie riftsqqkii qnalkhnqev iippgikftv
61 vtvkakparq ghnpatgepi qikakpehka vkiralkpvh dmln
SEQUENCE LISTING
<160> 4
<170> PatentIn version 3.3
<210> 1
<211> 142
<212> PRT
＜213〉P72 albumen
<400> 1
1 vsvegtsgpl lcnihdlhkp hqskpiltde ndtqrtcsht npkflsqhfp enshniqtag
61 kqditpitda tyldirrnvh yscngpqtpk yyqpplalwi klrfwfnenv nlaipsvsip
121 fgerfitikl asqkdlvnef pg
<210> 2
<211> 205
<212> PRT
＜213〉K205R albumen
<400> 2
1 mvepreqffq dllsavdqqm dtvkndikdi mkektsfmvs fenfierydt mekniqdlqn
61 kyeemaanlm tvmtdtkiql gaiiaqleil mingtplpak kttikeampl pssntnneqt
121 sppasgktse tpkknptnam fftrsewass ntfrekfltp eiqaildeqf anktgierlh
181 aeglymwrtq fsdeqkkmvk emmkk
<210> 3
<211> 186
<212> PRT
＜213〉P54 albumen
<400> 3
1 mdseffqpvy prhygeclss tptpsffsth mytiliaivv liiiiivliy lfssrkkkaa
61 aaieeediqf inpyqdqqwa gvppqpgiak pagattgsvs kpvmdrpitn npvmdrpvtn
121 hpvtdrlvtd klgmatgepa aasapahsae pyttvttqnt asqtmsaien lrqrstythk
181 dlensl
<210> 4
<211> 104
<212> PRT
＜213〉A104R albumen
<400> 4
1 mstkkkptit kqelyslvaa dtqlnkalie riftsqqkii qnalkhnqev iippgikftv
61 vtvkakparq ghnpatgepi qikakpehka vkiralkpvh dmln
Claims (5)
1. biological safety African swine fever antigen multiple-factor serum manufacture method, it is characterized in that, adopt four kinds of albumen of structural protein P72, K205R, P54, A104R of gene expression, through chemical purification, be coated with freund 's incomplete adjuvant, in three batches time muscle immunization experiment animal pig, gather Sanguis sus domestica after one month, separation of serum detects through serology, and packing is preserved.
2. biological safety African swine fever antigen multiple-factor serum manufacture method as claimed in claim 1, it is characterized in that: structural protein P72 Argine Monohydrochloride sequence is as follows:
1 vsvegtsgpl lcnihdlhkp hqskpiltde ndtqrtcsht npkflsqhfp enshniqtag
61 kqditpitda tyldirrnvh yscngpqtpk yyqpplalwi klrfwfnenv nlaipsvsip
121 fgerfitikl asqkdlvnef pg。
3. biological safety African swine fever antigen multiple-factor serum manufacture method as claimed in claim 1, it is characterized in that: structural protein K205R Argine Monohydrochloride sequence is as follows:
1 mvepreqffq dllsavdqqm dtvkndikdi mkektsfmvs fenfierydtmekniqdlqn
61 kyeemaanlm tvmtdtkiql gaiiaqleil mingtplpak kttikeamplpssntnneqt
121 sppasgktse tpkknptnam fftrsewass ntfrekfltp eiqaildeqfanktgierlh
181 aeglymwrtq fsdeqkkmvk emmkk。
4. biological safety African swine fever antigen multiple-factor serum manufacture method as claimed in claim 1, it is characterized in that: structural protein P54 Argine Monohydrochloride sequence is as follows:
1 mdseffqpvy prhygeclss tptpsffsth mytiliaivv liiiiivliy lfssrkkkaa
61 aaieeediqf inpyqdqqwa gvppqpgiak pagattgsvs kpvmdrpitn npvmdrpvtn
121 hpvtdrlvtd klgmatgepa aasapahsae pyttvttqnt asqtmsaienlrqrstythk
181 dlensl。
5. biological safety African swine fever antigen multiple-factor serum manufacture method as claimed in claim 1, it is characterized in that: structural protein A104R Argine Monohydrochloride sequence is as follows:
1 mstkkkptit kqelyslvaa dtqlnkalie riftsqqkii qnalkhnqeviippgikftv
61 vtvkakparq ghnpatgepi qikakpehka vkiralkpvh dmln。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103271218A CN102949720A (en) | 2012-09-06 | 2012-09-06 | Biosecurity multifactor blood serum production method for African swine fever |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103271218A CN102949720A (en) | 2012-09-06 | 2012-09-06 | Biosecurity multifactor blood serum production method for African swine fever |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102949720A true CN102949720A (en) | 2013-03-06 |
Family
ID=47759480
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012103271218A Pending CN102949720A (en) | 2012-09-06 | 2012-09-06 | Biosecurity multifactor blood serum production method for African swine fever |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102949720A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109810948A (en) * | 2019-01-18 | 2019-05-28 | 中国农业科学院兰州兽医研究所 | The hybridoma cell strain of anti-African swine fever virus K205R protein monoclonal antibody and its antibody of secretion |
| CN110235901A (en) * | 2019-07-10 | 2019-09-17 | 广州市德百顺电气科技有限公司 | A kind of thimerosal and its preparation facilities for prevention and control African swine fever |
| CN111363016A (en) * | 2020-03-30 | 2020-07-03 | 中国农业大学 | African swine fever immune antigen and preparation method and application thereof |
| CN112444626A (en) * | 2019-08-30 | 2021-03-05 | 洛阳普泰生物技术有限公司 | African swine fever virus antibody ELISA detection kit and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101833005A (en) * | 2010-04-23 | 2010-09-15 | 天津出入境检验检疫局动植物与食品检测中心 | Competitive ELISA (Enzyme-Linked Immuno Sorbent Assay) kit for detecting antibody of African swine fever virus and application thereof |
| CN102236017A (en) * | 2010-04-23 | 2011-11-09 | 陈文刚 | Indirect ELISA kit for detecting African swine fever virus antibody and application thereof |
-
2012
- 2012-09-06 CN CN2012103271218A patent/CN102949720A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101833005A (en) * | 2010-04-23 | 2010-09-15 | 天津出入境检验检疫局动植物与食品检测中心 | Competitive ELISA (Enzyme-Linked Immuno Sorbent Assay) kit for detecting antibody of African swine fever virus and application thereof |
| CN102236017A (en) * | 2010-04-23 | 2011-11-09 | 陈文刚 | Indirect ELISA kit for detecting African swine fever virus antibody and application thereof |
Non-Patent Citations (6)
| Title |
|---|
| ANA LUISA REIS 等: "Systematic analysis of longitudinal serological responses of pigs infected experimentally with African swine fever virus", 《JOURNAL OF GENERAL VIROLOGY》 * |
| BASTOS,A.D.S.等: "AAQ08042", 《GENBANK》 * |
| CHAPMAN,D.A.等: "CBW46714", 《GENBANK》 * |
| MISINZO,G.等: "ADD62407", 《GENBANK》 * |
| YANEZ,R.J.等: "NP_042728", 《GENBANK》 * |
| 蒋正军 等: "非洲猪瘟间接ELISA诊断试剂盒的研究", 《中国预防兽医学报》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109810948A (en) * | 2019-01-18 | 2019-05-28 | 中国农业科学院兰州兽医研究所 | The hybridoma cell strain of anti-African swine fever virus K205R protein monoclonal antibody and its antibody of secretion |
| CN109810948B (en) * | 2019-01-18 | 2021-09-17 | 中国农业科学院兰州兽医研究所 | Hybridoma cell strain of monoclonal antibody against African swine fever virus K205R protein and antibody secreted by hybridoma cell strain |
| CN110235901A (en) * | 2019-07-10 | 2019-09-17 | 广州市德百顺电气科技有限公司 | A kind of thimerosal and its preparation facilities for prevention and control African swine fever |
| CN112444626A (en) * | 2019-08-30 | 2021-03-05 | 洛阳普泰生物技术有限公司 | African swine fever virus antibody ELISA detection kit and preparation method thereof |
| CN112444626B (en) * | 2019-08-30 | 2023-09-12 | 洛阳普泰生物技术有限公司 | African swine fever virus antibody ELISA detection kit and preparation method thereof |
| CN111363016A (en) * | 2020-03-30 | 2020-07-03 | 中国农业大学 | African swine fever immune antigen and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102967703B (en) | A kind of biological safety African swine fever antigen multiple-factor serum for ELISA diagnosis | |
| Bello et al. | Diagnostic and vaccination approaches for Newcastle disease virus in poultry: The current and emerging perspectives | |
| Tao et al. | Bovine viral diarrhea virus (BVDV) infections in pigs | |
| CN103751774B (en) | The recombinant cell lines of stably express CSFV E 2 protein and in the application of preparing in subunit vaccine for swine fever and diagnostic reagent | |
| Farooq et al. | Characterization of naturally occurring, new and persistent subclinical foot‐and‐mouth disease virus infection in vaccinated Asian buffalo in Islamabad Capital Territory, Pakistan | |
| Wright | Serological and genetic characterisation of putative new serotypes of bluetongue virus and epizootic haemorrhagic disease virus isolated from an alpaca | |
| von Teichman et al. | In vivo cross-protection to African horse sickness Serotypes 5 and 9 after vaccination with Serotypes 8 and 6 | |
| CN103320535B (en) | Method for identifying wild strain and vaccine strain of hog cholera virus | |
| Ciacci-Zanella et al. | Influenza A virus infection in Brazilian swine herds following the introduction of pandemic 2009 H1N1 | |
| Paton et al. | Developing vaccines against foot-and-mouth disease and some other exotic viral diseases of livestock | |
| Qiang et al. | The effects of H9N2 influenza A on the immune system of broiler chickens in the Shandong Province | |
| CN102949720A (en) | Biosecurity multifactor blood serum production method for African swine fever | |
| CN107056898A (en) | 3 type of carp herpesviral, 1301 plants of ORF136 DNA recombinant expressions albumen, antibody and its application | |
| CN105671003A (en) | Infectious bronchitis low-virulent live vaccine YX10 D90 strain | |
| Reneer et al. | Broadly reactive H2 hemagglutinin vaccines elicit cross-reactive antibodies in ferrets preimmune to seasonal influenza a viruses | |
| CN102168088A (en) | T cell immunogen gene TI and applications thereof in foot-and-mouth disease protein subunit vaccine and inactivated vaccine | |
| CN106755087B (en) | Stablize the expression recombinant cell lines of CSFV E 2 protein, preparation method, using and swine fever virus subunit vaccine | |
| Kim et al. | Cross-protective immune responses elicited by a Korean variant of infectious bronchitis virus | |
| Graham et al. | Cultural characteristics of salmonid alphaviruses–influence of cell line and temperature | |
| CN108315306A (en) | One plant height fertility swine fever virus and its construction method | |
| CN102735807B (en) | Effect testing method for duck hemorrhagic ovaritis inactivated vaccine | |
| CN105695629B (en) | The primer and method of sldh gene I type and II type duck reovirus | |
| CN101833002A (en) | Antibody indirect immunofluorescence test method for distinguishing immune animal infected with influenza A virus | |
| CN103160620B (en) | High-efficient PCR (polymerase chain reaction) detection method of infectious hematopoietic necrosis virus | |
| Wu et al. | A meta-analysis and experiment assessing phage-based FMDV vaccine |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20130306 |