CN110894243B - A porcine blue ear virus chimeric antigen and colloidal gold immunochromatographic test strip for detecting porcine blue ear virus antibody - Google Patents
A porcine blue ear virus chimeric antigen and colloidal gold immunochromatographic test strip for detecting porcine blue ear virus antibody Download PDFInfo
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- CN110894243B CN110894243B CN201911300052.XA CN201911300052A CN110894243B CN 110894243 B CN110894243 B CN 110894243B CN 201911300052 A CN201911300052 A CN 201911300052A CN 110894243 B CN110894243 B CN 110894243B
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- respiratory syndrome
- porcine reproductive
- chimeric antigen
- test strip
- colloidal gold
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Abstract
The invention relates to the technical field of immunodetection, and discloses a porcine reproductive and respiratory syndrome virus chimeric antigen and a colloidal gold immunochromatographic test strip for detecting a porcine reproductive and respiratory syndrome virus antibody. The chimeric antigen sequence is shown in SEQ ID NO. 1. The invention provides a novel porcine reproductive and respiratory syndrome virus chimeric antigen, which has high specificity, repeatability and accuracy when being used as a detection antigen of a porcine reproductive and respiratory syndrome virus antibody, can be applied to related immunochromatography detection products, has intuitive and visual results, can quickly and specifically detect the porcine reproductive and respiratory syndrome virus antibody in serum, has high practical value and is convenient for basic popularization and use.
Description
Technical Field
The invention relates to the technical field of immunodetection, and particularly relates to a porcine reproductive and respiratory syndrome virus chimeric antigen and a colloidal gold immunochromatographic test strip for detecting a porcine reproductive and respiratory syndrome virus antibody.
Background
Porcine Reproductive and Respiratory Syndrome (PRRS), caused by PRRS, is a contagious disease mainly characterized by high fever, high morbidity and mortality, adult porcine reproductive disorders, abortion and stillbirth, and respiratory abnormality of piglets. Because of the serious threat to the swine industry, the vaccine is registered as a B-type infectious disease by the International Bureau of zooepidemics.
The detection method of the porcine reproductive and respiratory syndrome virus antibody has various methods, and the laboratory diagnosis technologies established and applied at present comprise virus separation and identification, immunoperoxidase monolayer test, indirect immunofluorescence test, indirect enzyme-linked immunosorbent test, serum neutralization test and the like. The virus separation and identification are mostly used for the confirmation of acute cases and the determination of new epidemic areas, and the immunoperoxidase monolayer test, the indirect immunofluorescence test and the indirect enzyme-linked immunosorbent test are similar to the specificity of virus antibody detection, have high requirements on technical conditions and expensive equipment, so the application of the virus antibody detection in a basic layer is limited. Neutralizing antibodies appear late and are not suitable for early diagnostic applications.
At present, the indirect enzyme-linked immunosorbent assay mostly adopts a virus-coated plate or an N protein-coated plate, so that the virus is easy to disperse, and the detection has the risk of missing detection. Aiming at the epidemic situation and the prevention and control situation of the porcine reproductive and respiratory syndrome, the antibody diagnosis of the porcine reproductive and respiratory syndrome requires high specificity and strong sensitivity, and also requires simple and convenient operation, simple and clear results and easy use by primary epidemic prevention units, farms and farmers.
Disclosure of Invention
In view of the above, the present invention aims to provide a porcine reproductive and respiratory syndrome virus chimeric antigen, which has high specificity, repeatability and accuracy when applied to detection of porcine reproductive and respiratory syndrome antibodies;
the invention also aims to provide the related application of the porcine reproductive and respiratory syndrome virus chimeric antigen in the preparation of an immunochromatography detection product.
In order to achieve the above purpose, the invention provides the following technical scheme:
a porcine reproductive and respiratory syndrome virus chimeric antigen has a sequence shown in SEQ ID NO 1.
The invention adopts bioinformatics technology to carry out antigenicity and conservative analysis on structural protein N protein and GP5 protein of different pedigree strains of the porcine reproductive and respiratory syndrome virus, designs and synthesizes a porcine reproductive and respiratory syndrome virus specific chimeric antigen NG gene (shown in SEQ ID NO: 2), and then obtains the antigen through a recombinant expression mode.
The antigen is expressed by recombination as follows:
inserting the porcine reproductive and respiratory syndrome virus specific chimeric antigen NG gene between Hind III and NotI enzyme cutting sites of the vector pGAPZ, keeping other sequences of the pGAPZ unchanged to obtain a pGAPZ-NG gene recombinant expression vector, and then converting the pGAPZ-NG gene recombinant expression vector into pichia pastoris expression engineering bacteria SMD1168 to insert the pichia pastoris expression engineering bacteria SMD1168 into a pichia pastoris genome to stably express a target protein.
In the specific implementation mode of the invention, the antigen is used for preparing the colloidal gold immunochromatographic test strip, and the specific test result shows that the cross test is carried out on the known O-type positive serum of the pig foot-and-mouth disease, the positive serum of the pig circovirus II, the positive serum of the pig rotavirus, the positive serum of the classical swine fever virus and the positive serum of the pig blue-ear virus. Only the positive serum sample of the porcine reproductive and respiratory syndrome virus is detected to be colored and show positive reaction, while when the detection line is reacted with other several kinds of serum, the detection line is not colored and shows negative reaction, which indicates that the antigen has high specificity when being used for detecting the porcine reproductive and respiratory syndrome virus antibody.
Meanwhile, 5 verified porcine reproductive and respiratory syndrome virus antibody positive serums and 5 verified porcine reproductive and respiratory syndrome virus antibody negative serums are respectively detected, 5 replicates are conducted on each serum sample in parallel, and the result shows that the detection results of the batch repeatability test and the batch repeatability test are consistent, so that the antigen has better repeatability and accuracy when used for detecting the porcine reproductive and respiratory syndrome virus antibody.
In addition, 141 known clinical serum samples were tested with the same lot of colloidal gold immunochromatographic test strips, and the results were observed and compared with the data of the ELISA test kit coated with N-protein. ELISA detection data show that 141 serum samples are 107 positive samples, 34 negative samples, 1 false positive sample, 5 false negative samples, 95.5 percent of positive coincidence rate (106/111), 96.7 percent of negative coincidence rate (29/30) and 95.7 percent of total coincidence rate (135/141); the test strip of the invention has 109 positive samples, 32 negative samples, 2 false negative numbers, 98 percent (109/111) positive coincidence rate, 100 percent negative coincidence rate and 98.6 percent (139/141) total coincidence rate, which shows that the chimeric antigen has good accuracy when being used for detecting the porcine reproductive and respiratory syndrome virus antibody.
Based on the test result, the invention provides the application of the chimeric antigen in preparing an immunochromatography product for detecting the porcine reproductive and respiratory syndrome virus antibody. Preferably, the immunochromatographic product is a colloidal gold immunochromatographic test strip.
According to the application, the invention provides a colloidal gold immunochromatographic test strip for detecting a porcine reproductive and respiratory syndrome virus antibody, which takes a protein with a sequence shown in SEQ ID NO. 1 as a detection antigen.
Preferably, the test strip comprises a backing, and a sample pad, a marker pad, a nitrocellulose membrane and a water absorption pad which are sequentially stuck on the backing, wherein the nitrocellulose membrane is scribed with a detection line of the porcine reproductive and respiratory syndrome virus chimeric antigen coated with a sequence shown in SEQ ID NO. 1. The spraying amount of the porcine reproductive and respiratory syndrome virus chimeric antigen on the detection line is 1ul/cm, and the working concentration of the porcine reproductive and respiratory syndrome chimeric antigen is 3 mg/ml.
Preferably, a quality control line coated with the rabbit anti-pig IgG antibody is scribed on the nitrocellulose membrane, and the spraying amount of the rabbit anti-pig IgG antibody on the quality control line is 1 ul/cm; the working concentration of the rabbit anti-bovine antibody is 2 mg/ml.
Preferably, the label pad is sprayed with the compound of staphylococcus aureus protein a (spa) labeled with the colloidal gold particles, and the spraying amount of the compound of staphylococcus aureus protein a (spa) labeled with the colloidal gold particles on the label pad is 8 uL/cm. In a specific embodiment of the present invention, the labeling ratio content of the colloidal gold to the SPA in the colloidal gold labeling process is: 2ml of colloidal gold: 10ug of SPA.
Preferably, the backing is a polyethylene backing.
According to the technical scheme, the novel porcine reproductive and respiratory syndrome chimeric antigen provided by the invention has extremely high specificity, repeatability and accuracy when being used as a detection antigen of a porcine reproductive and respiratory syndrome antibody, can be applied to related immunochromatography detection products, can be visually judged by naked eyes, can be used for quickly and specifically detecting the porcine reproductive and respiratory syndrome antibody in serum, has higher practical value, and is convenient for basic popularization and use.
Drawings
FIG. 1 is a schematic diagram of a colloidal gold immunochromatographic test strip;
FIG. 2 shows the specificity analysis of the colloidal gold immunochromatographic test strip; wherein, 1 represents a porcine reproductive and respiratory syndrome virus positive serum; 2 represents the O-type positive serum of the pig foot-and-mouth disease; 3 represents porcine circovirus type II positive serum; 4 represents porcine rotavirus positive serum; and 5 represents swine fever virus positive serum.
Detailed Description
The invention discloses a porcine reproductive and respiratory syndrome virus chimeric antigen and a colloidal gold immunochromatographic test strip for detecting a porcine reproductive and respiratory syndrome antibody. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the chimeric porcine reproductive and respiratory syndrome virus antigen and the colloidal gold immunochromatographic strip of the present invention have been described in terms of preferred embodiments, it will be apparent to those skilled in the art that modifications, or appropriate alterations and combinations thereof can be made to the chimeric porcine reproductive and respiratory syndrome antigen and the colloidal gold immunochromatographic strip of the present invention to achieve and utilize the techniques of the present invention without departing from the spirit and scope of the present invention.
When the detected sample contains the porcine reproductive and respiratory syndrome virus antibody, the colloidal gold marked SPA and the porcine reproductive and respiratory syndrome virus antibody in the sample form a compound which moves in a chromatography mode and is captured by the porcine reproductive and respiratory syndrome virus chimeric antigen NG sprayed on a nitrocellulose membrane at a T line position, the T line forms a gold marked SPA-porcine reproductive and respiratory syndrome antibody-porcine reproductive and respiratory syndrome chimeric antigen NG compound, the unbound gold marked SPA-porcine IgG continues to move to a C line and is captured by the rabbit anti-porcine IgG antibody to form a gold marked SPA-porcine IgG-rabbit anti-porcine IgG antibody compound, and at the moment, the T line and the C line both show red visible to naked eyes. When the sample to be detected does not contain the porcine reproductive and respiratory syndrome virus antibody, the gold-labeled SPA-porcine IgG compound does not react with the porcine reproductive and respiratory syndrome chimeric antigen NG on the T line, the gold-labeled SPA-antibody compound continuously moves to the C line and is captured by the rabbit anti-porcine IgG to form the gold-labeled SPA-antibody-rabbit anti-porcine IgG compound, the T line does not develop color, and the C line shows red visible to naked eyes. And (4) judging a result: the T line is not colored, the C line is colored red, the detection sample is negative, the T line and the C line are both colored red, the detection sample is positive, and if the C line and the T line are not colored, the test strip is invalid.
The present invention provides a porcine reproductive and respiratory syndrome virus chimeric antigen and a colloidal gold immunochromatographic test strip for detecting porcine reproductive and respiratory syndrome virus antibodies.
Example 1: recombinant expression of the chimeric antigens of the invention
1. Construction of porcine reproductive and respiratory syndrome virus chimeric antigen gene recombinant expression plasmid pGAPZ-NG
The different pedigree strain structural protein N protein and GP5 protein of the porcine reproductive and respiratory syndrome virus are subjected to antigenicity and conservative analysis by using a bioinformatics technology, a porcine reproductive and respiratory syndrome virus specific chimeric antigen NG gene is designed and synthesized and optimized, and a porcine reproductive and respiratory syndrome chimeric antigen gene sequence (shown in SEQ ID NO: 2) is synthesized by using a chemical synthesis method. The pGAPZ vector was double digested with HindIII and NotI in a 50 uL: HindIII and NotI each 1uL, vector fragment 20uL, 10 Xdigestion buffer 5uL, ddH2O 23uL, digestion products through 1% agarose electrophoresis and gel recovery kit recovery. Then, the enzyme-cut vector is connected with the NG gene sequence respectively to construct a 10 mu L connection system: 5.5 μ L of NG gene fragment, 2.5 μ L of pGAPZ vector, 0.5 μ L of T4DNA ligase, and 1 μ L of T4DNA ligation buffer. Tapping the tube wall, mixing uniformly, connecting for 1h at room temperature, converting DH5 alpha competent cells of the connecting product by a conventional chemical method, and identifying positive clones by a PCR method. The positive clones which are correctly identified by PCR are sequenced, and the plasmid which is positive in sequencing is named pGAPZ-NG.
2. Recombinant expression of chimeric antigen proteins
The positive clone plasmid from the extraction screening was transformed into SMD1168 by chemical method. NG gene insertion was verified by colony PCR. By increasing the concentration of Zeocin antibiotic, high copy transformant is screened out. The positive single clone Pichia pastoris colony is selected and inoculated in YPD liquid culture medium and cultured overnight at 30 ℃. And (3) taking the overnight cultured bacterial liquid to perform 100-fold amplification culture in the MGY liquid culture medium, and culturing at 30 ℃ for 60 hours to obtain the required protein.
3. Purification of chimeric antigen proteins
Taking 1L of Pichia pastoris culture, centrifuging at 6000rpm for 10min, resuspending the precipitate with NTA-0buffer, crushing with a high pressure homogenizer, and centrifuging at 18000rpm for 30min at 4 ℃. The supernatant was filtered through a 0.45 μm filter and purified.
The method uses a nickel column for affinity purification of the fusion protein and comprises the following main operation steps:
a. the column was equilibrated with 20mL of NTA-0buffer, and the centrifuged supernatant was slowly added to the Ni column using a syringe.
b. The mixture was washed with 20mL of NTA-0buffer, and the flow rate was controlled to be about 1 mL/min.
c. Sequentially eluting with 20mL NTA-50, NTA-100 and NTA-500buffer at flow rate of about 1 mL/min; the eluted peak fractions were collected.
d. The collected samples were subjected to SDS-PAGE analysis.
Example 2: colloidal gold immunochromatography detection test strip for preparing porcine reproductive and respiratory syndrome virus antibody
1. Preparation of gold-labeled conjugates of SPA labeled with gold particles
Taking 2ml of 40nm colloidal gold particles, adjusting the pH value to 6.8 by using 0.1mol/L potassium carbonate adjusting solution, adding 10ug of staphylococcus aureus protein A (SPA), quickly and uniformly mixing the gold particles and the staphylococcus aureus protein A (SPA), uniformly mixing the gold particles and the staphylococcus aureus protein A at room temperature on a 3D rotating instrument for 30min, then adding BSA (bovine serum albumin) with the final concentration of 1 percent, uniformly mixing the BSA at the 3D rotating instrument for 30min, centrifuging a gold standard solution at the temperature of 4 ℃ and 12000r/min for 10min, carefully removing a supernatant, washing a precipitate for 2 times by using 0.01M PBS (phosphate buffer solution), centrifuging the obtained precipitate again to obtain a purified SPA-labeled compound, re-suspending the prepared colloidal gold-labeled SPA by using 0.01 MP.
2. Nitrocellulose membrane detection line and quality control line spraying and colloidal gold combined pad preparation
The recombinant porcine reproductive and respiratory syndrome chimeric antigen NG is diluted to 3mg/ml by 0.01MPBS and evenly scribed on the T line position of a nitrocellulose membrane by 1 ul/cm. The rabbit anti-pig IgG antibody was diluted to 2mg/ml with 0.01MPBS, evenly scratched at 1ul/cm on the C line of nitrocellulose membrane, and then dried overnight at 37 ℃ for use. SPA-gold colloidal complex was sprayed evenly onto the marker pad at 8uL/cm and dried overnight at 37 ℃ for use.
3. Assembly of test strips
Sequentially adhering a sample pad, a marking pad sprayed with an SPA gold-labeled conjugate, a nitrocellulose membrane sprayed with a porcine reproductive and respiratory syndrome virus chimeric antigen NG as a detection line and a rabbit anti-porcine IgG as a quality control line, and an absorption pad on a polyethylene back lining. The assembly structure is that the nitrocellulose membrane is arranged on the polyethylene back lining, the sample pad is flatly attached to the left side of the nitrocellulose membrane, the absorption pad is flatly attached to the right side of the nitrocellulose membrane, the colloidal gold combination pad is flatly attached between the sample pad and the nitrocellulose membrane, one end of the colloidal gold combination pad is pressed below the sample pad, and the other end of the colloidal gold combination pad is covered on the nitrocellulose membrane. The assembled test strip is packaged in vacuum and stored at normal temperature; the schematic view is shown in FIG. 1.
4. Application of colloidal gold immunochromatographic test strip for detecting porcine reproductive and respiratory syndrome virus antibody
150ul of sample to be tested is dripped on the test paper sample pad, the result is observed after 10min, and meanwhile, a known positive serum sample and a known negative serum are respectively taken as positive control and negative control. And (4) judging a result: when a detection sample is added on the sample pad, if the detection line and the quality control line are both red, the detection line is positive to the porcine reproductive and respiratory syndrome virus antibody; if the detection line does not develop color and the quality control line develops red, the detection line is negative to the porcine reproductive and respiratory syndrome virus antibody; if the quality control line does not develop color, the test paper is invalid.
Example 3: colloidal gold immunochromatographic test strip specificity test
Known swine foot-and-mouth disease O-type positive serum, swine circovirus type II positive serum, swine rotavirus positive serum, swine fever virus positive serum and swine blue ear virus positive serum are subjected to cross tests, and the results are shown in figure 2.
The result of figure 2 shows that only the positive serum sample of the porcine reproductive and respiratory syndrome virus shows the detection line is colored and shows a positive reaction, but when the detection line is reacted with other several kinds of serum, the detection line is not colored and shows a negative reaction, which indicates that the chimeric antigen has high specificity when being used for detecting the antibody of the porcine reproductive and respiratory syndrome virus.
Example 4: test paper strip repeatability and accuracy test of colloidal gold immunochromatography
5 parts of verified porcine reproductive and respiratory syndrome virus antibody positive serum and 5 parts of verified porcine reproductive and respiratory syndrome virus antibody negative serum are respectively detected by the immunochromatographic colloidal gold test paper of different batches, 5 repetitions are carried out on each serum sample in parallel, and the results show that the detection results of the batch repeatability test and the batch repeatability test are consistent and are consistent with the virus neutralization test result, which shows that the colloidal gold immunochromatographic test paper strip has good repeatability and accuracy.
Example 5: compliance test of colloidal gold immunochromatographic test strip
141 parts of known clinical serum samples are respectively detected by the same batch of colloidal gold immunochromatographic test strips, and the results are observed and compared with the detection data of the ELISA detection kit coated by the N protein. ELISA detection data show that 141 serum samples are 107 positive samples, 34 negative samples, 1 false positive sample, 5 false negative samples, 95.5 percent of positive coincidence rate (106/111), 96.7 percent of negative coincidence rate (29/30) and 95.7 percent of total coincidence rate (135/141); the test strip of the invention has 109 positive samples, 32 negative samples, 2 false negative numbers, 98 percent (109/111) positive coincidence rate, 100 percent negative coincidence rate and 98.6 percent (139/141) total coincidence rate, which shows that the chimeric antigen has good accuracy when being used for detecting the porcine reproductive and respiratory syndrome virus antibody.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Sequence listing
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Citations (25)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998050426A1 (en) * | 1997-05-06 | 1998-11-12 | Stichting Dienst Landbouwkundig Onderzoek | Prrsv antigenic sites identifying peptide sequences of prrs virus for use in vaccines or diagnostic assays |
| CN101184997A (en) * | 2005-02-25 | 2008-05-21 | 艾德克斯实验室公司 | Peptides for detecting antibodies against porcine reproductive and respiratory syndrome virus |
| CN101216488A (en) * | 2008-01-10 | 2008-07-09 | 中国农业科学院哈尔滨兽医研究所 | Test strip for detecting porcine reproductive and respiratory syndrome antibody and preparation method thereof |
| CN101236205A (en) * | 2008-01-23 | 2008-08-06 | 山东省农业科学院畜牧兽医研究所 | European and America type pig breeding and syndrome virus antibody identification and diagnosis reagent kit preparation method |
| CN101696399A (en) * | 2009-10-22 | 2010-04-21 | 中国农业科学院哈尔滨兽医研究所 | Porcine reproductive and respiratory syndrome virus M protein CTL cell epitopes and application thereof |
| CN202614765U (en) * | 2011-12-28 | 2012-12-19 | 滨州职业学院 | Porcine reproductive and respiratory syndrome virus PPA-ELISA (enzyme-linked immuno sorbent assay) antibody detection kit |
| CN103275193A (en) * | 2013-05-28 | 2013-09-04 | 福建省农业科学院畜牧兽医研究所 | Indirect ELISA (enzyme-linked immunosorbent assay) method for detecting PRRSV (porcine reproductive and respiratory syndrome virus) antibody through tandem repeat expression of GP5 dominant antigen epitopes |
| CN103641921A (en) * | 2013-12-05 | 2014-03-19 | 中国人民解放军第三军医大学第一附属医院 | Multi-epitope fusion antigen and kit for detecting porcine reproductive and respiratory syndrome virus serum antibody |
| CN103648527A (en) * | 2011-12-30 | 2014-03-19 | 美国联合生物医学公司 | Synthetic peptide-based marker vaccine and diagnostic system for effective control of porcine reproductive and respiratory syndrome (PRRS) |
| CN103995136A (en) * | 2014-06-06 | 2014-08-20 | 武汉中博生物股份有限公司 | Rapid colloidal gold detection test strip for pig porcine reproductive and respiratory syndrome virus |
| KR101430223B1 (en) * | 2012-10-09 | 2014-09-25 | 전북대학교산학협력단 | Porcine Reproductive and Respiratory Syndrome Virus having enhanced immune-inducing property |
| CN203949923U (en) * | 2013-12-17 | 2014-11-19 | 深圳市宝安康生物技术有限公司 | A kind of rapid detection card that detects pig blue-ear disease poison antibody |
| CN204008662U (en) * | 2014-04-24 | 2014-12-10 | 北京维德维康生物技术有限公司 | Detect the ELISA kit of PRRS virus antibody |
| CN104330572A (en) * | 2014-10-27 | 2015-02-04 | 苏州市吴江区畜牧兽医站 | Kit for indirect ELISA (Enzyme-Linked Immunosorbent Assay) detection of IgG or IgA antibodies of PRRSV (Porcine reproductive and respiratory syndrome virus) of pigs |
| CN104459160A (en) * | 2014-12-30 | 2015-03-25 | 洛阳普莱柯万泰生物技术有限公司 | Kit and preparation method and application method thereof |
| WO2015092058A1 (en) * | 2013-12-20 | 2015-06-25 | Boehringer Ingelheim Vetmedica Gmbh | Prrs virus variant, european prrs virus cdna clone, and uses thereof |
| CN105424928A (en) * | 2015-11-11 | 2016-03-23 | 山西隆克尔生物制药有限公司 | Immunochromatography strip detecting porcine reproductive and respiratory syndrome virus antibodies and preparation method and application thereof |
| CN105606820A (en) * | 2016-01-21 | 2016-05-25 | 成都微瑞生物科技有限公司 | Porcine reproductive and respiratory syndrome antibody detecting card and preparation method thereof |
| CN205374469U (en) * | 2016-01-21 | 2016-07-06 | 成都微瑞生物科技有限公司 | Blue ear virus antibody detection card of pig |
| CN106075422A (en) * | 2005-11-29 | 2016-11-09 | 衣阿华州立大学研究基金公司 | Qualification of protective antigenic determinants of porcine reproductive and respiratory syndrome virus (PRRSV) and application thereof |
| WO2017096468A1 (en) * | 2015-12-10 | 2017-06-15 | Aerovirus Technologies Inc. | Aptamers, nucleic acid molecules, polynucleotides, synthetic antibodies compositions for detecting prrs viruses and treating prrs virus infection |
| CN108508210A (en) * | 2018-02-13 | 2018-09-07 | 中国农业科学院兰州兽医研究所 | The prokaryotic soluble expression method and indirect ELISA antibody detection method of PRRSV N proteins |
| CN108559754A (en) * | 2018-02-13 | 2018-09-21 | 南京农业大学 | A kind of recombinant protein of porcine reproductive and respiratory syndrome virus GP5 genetic modifications and its application |
| CN108872602A (en) * | 2018-07-10 | 2018-11-23 | 安徽九川生物科技有限公司 | A kind of immune Magnetism particulate immuno chemistry luminescence method immue quantitative detection reagent box and its application method of porcine reproductive and respiratory syndrome antibody |
| WO2019004811A1 (en) * | 2017-06-29 | 2019-01-03 | Centro De Investigación Y Asistencia En Tecnología Y Diseño Del Estado De Jalisco A.C. | Chimeric protein for the prevention and diagnosis of porcine respiratory and reproductive syndrome (prrs) |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040014028A1 (en) * | 2002-07-19 | 2004-01-22 | Wolf Biotech | Method for determination of protective epitopes for vaccination, diagnosis and vaccine quality control |
| US7241582B2 (en) * | 2005-07-05 | 2007-07-10 | Mj Biologics, Inc. | Diagnostic test kits |
-
2019
- 2019-12-16 CN CN201911300052.XA patent/CN110894243B/en active Active
Patent Citations (26)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998050426A1 (en) * | 1997-05-06 | 1998-11-12 | Stichting Dienst Landbouwkundig Onderzoek | Prrsv antigenic sites identifying peptide sequences of prrs virus for use in vaccines or diagnostic assays |
| UA73915C2 (en) * | 1997-05-06 | 2005-10-17 | Стіхтінг Дінст Ландбаувкюндіг Ондерзук | Antigenic sites of prrsv defining sequences of proteins of ppcc virus being suitable for use in vaccines or in diagnostics |
| CN101184997A (en) * | 2005-02-25 | 2008-05-21 | 艾德克斯实验室公司 | Peptides for detecting antibodies against porcine reproductive and respiratory syndrome virus |
| CN106075422A (en) * | 2005-11-29 | 2016-11-09 | 衣阿华州立大学研究基金公司 | Qualification of protective antigenic determinants of porcine reproductive and respiratory syndrome virus (PRRSV) and application thereof |
| CN101216488A (en) * | 2008-01-10 | 2008-07-09 | 中国农业科学院哈尔滨兽医研究所 | Test strip for detecting porcine reproductive and respiratory syndrome antibody and preparation method thereof |
| CN101236205A (en) * | 2008-01-23 | 2008-08-06 | 山东省农业科学院畜牧兽医研究所 | European and America type pig breeding and syndrome virus antibody identification and diagnosis reagent kit preparation method |
| CN101696399A (en) * | 2009-10-22 | 2010-04-21 | 中国农业科学院哈尔滨兽医研究所 | Porcine reproductive and respiratory syndrome virus M protein CTL cell epitopes and application thereof |
| CN202614765U (en) * | 2011-12-28 | 2012-12-19 | 滨州职业学院 | Porcine reproductive and respiratory syndrome virus PPA-ELISA (enzyme-linked immuno sorbent assay) antibody detection kit |
| CN103648527A (en) * | 2011-12-30 | 2014-03-19 | 美国联合生物医学公司 | Synthetic peptide-based marker vaccine and diagnostic system for effective control of porcine reproductive and respiratory syndrome (PRRS) |
| KR101430223B1 (en) * | 2012-10-09 | 2014-09-25 | 전북대학교산학협력단 | Porcine Reproductive and Respiratory Syndrome Virus having enhanced immune-inducing property |
| CN103275193A (en) * | 2013-05-28 | 2013-09-04 | 福建省农业科学院畜牧兽医研究所 | Indirect ELISA (enzyme-linked immunosorbent assay) method for detecting PRRSV (porcine reproductive and respiratory syndrome virus) antibody through tandem repeat expression of GP5 dominant antigen epitopes |
| CN103641921A (en) * | 2013-12-05 | 2014-03-19 | 中国人民解放军第三军医大学第一附属医院 | Multi-epitope fusion antigen and kit for detecting porcine reproductive and respiratory syndrome virus serum antibody |
| CN203949923U (en) * | 2013-12-17 | 2014-11-19 | 深圳市宝安康生物技术有限公司 | A kind of rapid detection card that detects pig blue-ear disease poison antibody |
| WO2015092058A1 (en) * | 2013-12-20 | 2015-06-25 | Boehringer Ingelheim Vetmedica Gmbh | Prrs virus variant, european prrs virus cdna clone, and uses thereof |
| CN204008662U (en) * | 2014-04-24 | 2014-12-10 | 北京维德维康生物技术有限公司 | Detect the ELISA kit of PRRS virus antibody |
| CN103995136A (en) * | 2014-06-06 | 2014-08-20 | 武汉中博生物股份有限公司 | Rapid colloidal gold detection test strip for pig porcine reproductive and respiratory syndrome virus |
| CN104330572A (en) * | 2014-10-27 | 2015-02-04 | 苏州市吴江区畜牧兽医站 | Kit for indirect ELISA (Enzyme-Linked Immunosorbent Assay) detection of IgG or IgA antibodies of PRRSV (Porcine reproductive and respiratory syndrome virus) of pigs |
| CN104459160A (en) * | 2014-12-30 | 2015-03-25 | 洛阳普莱柯万泰生物技术有限公司 | Kit and preparation method and application method thereof |
| CN105424928A (en) * | 2015-11-11 | 2016-03-23 | 山西隆克尔生物制药有限公司 | Immunochromatography strip detecting porcine reproductive and respiratory syndrome virus antibodies and preparation method and application thereof |
| WO2017096468A1 (en) * | 2015-12-10 | 2017-06-15 | Aerovirus Technologies Inc. | Aptamers, nucleic acid molecules, polynucleotides, synthetic antibodies compositions for detecting prrs viruses and treating prrs virus infection |
| CN205374469U (en) * | 2016-01-21 | 2016-07-06 | 成都微瑞生物科技有限公司 | Blue ear virus antibody detection card of pig |
| CN105606820A (en) * | 2016-01-21 | 2016-05-25 | 成都微瑞生物科技有限公司 | Porcine reproductive and respiratory syndrome antibody detecting card and preparation method thereof |
| WO2019004811A1 (en) * | 2017-06-29 | 2019-01-03 | Centro De Investigación Y Asistencia En Tecnología Y Diseño Del Estado De Jalisco A.C. | Chimeric protein for the prevention and diagnosis of porcine respiratory and reproductive syndrome (prrs) |
| CN108508210A (en) * | 2018-02-13 | 2018-09-07 | 中国农业科学院兰州兽医研究所 | The prokaryotic soluble expression method and indirect ELISA antibody detection method of PRRSV N proteins |
| CN108559754A (en) * | 2018-02-13 | 2018-09-21 | 南京农业大学 | A kind of recombinant protein of porcine reproductive and respiratory syndrome virus GP5 genetic modifications and its application |
| CN108872602A (en) * | 2018-07-10 | 2018-11-23 | 安徽九川生物科技有限公司 | A kind of immune Magnetism particulate immuno chemistry luminescence method immue quantitative detection reagent box and its application method of porcine reproductive and respiratory syndrome antibody |
Non-Patent Citations (3)
| Title |
|---|
| AAK27366.1;Kang,S.Y等;《GenBank》;20010328;ORIGIN * |
| AEW90284.1;Hu,H.;《GenBank》;20111228;ORIGIN * |
| NP_047413.1;Allende，R.等;《GenBank》;20180813;ORIGIN * |
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