CN104569428B - A kind of enterovirns type 71 killed vaccine antigen enzyme-linked immunologic detecting kit - Google Patents
A kind of enterovirns type 71 killed vaccine antigen enzyme-linked immunologic detecting kit Download PDFInfo
- Publication number
- CN104569428B CN104569428B CN201410839523.5A CN201410839523A CN104569428B CN 104569428 B CN104569428 B CN 104569428B CN 201410839523 A CN201410839523 A CN 201410839523A CN 104569428 B CN104569428 B CN 104569428B
- Authority
- CN
- China
- Prior art keywords
- antibody
- enterovirns type
- enzyme
- solution
- vaccine antigen
- 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.)
- Active
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biotechnology (AREA)
- Analytical Chemistry (AREA)
- Cell Biology (AREA)
- Pathology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Peptides Or Proteins (AREA)
Abstract
The present invention relates to a kind of enterovirns type 71 killed vaccine antigen enzyme-linked immunologic detecting kit, described detection kit comprises: pre-coated anti-enterovirns type 71 polyclonal antibody ELISA Plate, sample diluting liquid, second antibody, enzyme labelled antibody, concentrated cleaning solution, enzyme substrate solution and stop buffer.ELISA Plate is wrapped by the polyclonal antibody prepared as immunogenic by recombination enterovirus 71 type structural proteins 1 in advance, and second antibody is the monoclonal antibody prepared as immunogenic by keyhole blood blue carrier protein couplet peptide sequence FGEHKQEKDL; Enzyme labelled antibody is the goat anti-mouse immune globulin antibody of horseradish peroxidase-labeled; Antigen standard is placed in box.The present invention, when measuring EV71 viral inactivation vaccine antigen titre, has good sensitivity, has linear preferably, linearly dependent coefficient r within the scope of 5.9-750ng/ml
2be greater than 0.99.Kit provided by the present invention is the detection kit that a species specificity is good, quantitatively accurate, highly sensitive, reproducible, measure EV71 viral inactivation vaccine antigen titre simply and easily.
Description
Technical field
The invention belongs to biological technical field, especially for enterovirns type 71 (EV71) killed vaccine antigen enzyme-linked immunologic detecting kit.
Background technology
In recent years, hand-foot-mouth disease wreaks havoc China's most area, has brought huge harm.A large amount of epidemiological studies shows, enterovirns type 71 is one of main pathogens causing hand-foot-and-mouth disease.EV71 belongs to Picornaviridae, enterovirus genus member.Catching of this virus, the multiple infant being born in less than 5 years old, can cause the bleb at the position such as hand, foot, oral cavity, and few patients can cause the complication such as myocarditis, pulmonary edema, AME, has higher disabling and fatal rate.Therefore develop the propagation that safe and effective EV71 vaccine carrys out prevention and control hand-foot-mouth disease, have very important significance.At present, commercially yet there are no relevant vaccine, and mainly comprise formalin-killed vaccine, viruslike particle (VLP) vaccine and nucleic acid vaccine etc. at the vaccine of research and development, wherein the fastest with the progress of formalin-killed vaccine again.
In the development process of EV71 inactivated vaccine, all need the detection carrying out EV71 viral antigen, although EV71 virus can produce cytopathy (CPE), cultured cell can be organized to detect the mensuration of carrying out EV71 antigenic content by virus infections.But the method cycle is longer, often need time a couple of days just can obtain result, and because CPE can only detect the virus of infection activity, make it correctly cannot detect antigenic content in inactivated vaccine, as vacuolating virus, activated virus etc., therefore cannot carry out effective, instant, correct monitoring in the production run of vaccine.And adopt euzymelinked immunosorbent assay (ELISA) to detect EV71 antigen, often because the reason of antibody sources affects to testing result, especially in vaccine, the non-specific adsorption of impurity causes great interference to testing result.
Summary of the invention
The object of the invention is to utilize two kinds of antibody with high specific to set up DASELISA immunization (ELISA), there is provided a species specificity good, highly sensitive, easy to use, detect enterovirns type 71 killed vaccine antigen detection kit fast, can be applied to enterovirns type 71 inactivated vaccine produce and quality arbitration.
The present invention relates to a kind of enterovirns type 71 killed vaccine antigen enzyme-linked immunologic detecting kit, described detection kit comprises: pre-coated anti-enterovirns type 71 polyclonal antibody ELISA Plate, sample diluting liquid, second antibody, enzyme labelled antibody, concentrated cleaning solution, enzyme substrate solution and stop buffer, ELISA Plate is wrapped by the polyclonal antibody prepared as immunogenic by recombination enterovirus 71 type structural proteins 1 in advance, and second antibody is the monoclonal antibody prepared as immunogenic by keyhole blood blue carrier protein couplet peptide sequence FGEHKQEKDL; Enzyme labelled antibody is the goat anti-mouse immune globulin antibody of horseradish peroxidase-labeled, and antigen standard is placed in box.Resist with the enterovirns type 71 that enterovirns type 71 capsid structural protein (VP1) recombinant antigen of Bacillus coli expression obtains as immunogenic more, enterovirns type 71 inactivated vaccine is detected there is better specificity, greatly can avoid the nonspecific reaction caused by vaccine cultured cell matrix, thus improve the accuracy detected.And using the mouse monoclonal antibody that enterovirns type 71 specific polypeptide FGEHKQEKDL obtains as immunogenic, because of specificity and the stronger immunogenicity thereof in polypeptide site used, make prepared monoclonal antibody have good specificity.The present invention, when measuring EV71 viral inactivation vaccine antigen titre, has good sensitivity, has linear preferably, linearly dependent coefficient r within the scope of 5.9-750ng/ml
2be greater than 0.99.Kit provided by the present invention is the detection kit that a species specificity is good, quantitatively accurate, highly sensitive, reproducible, measure EV71 viral inactivation vaccine antigen titre simply and easily.
The present invention is completed by following technical proposal: a kind of enterovirns type 71 killed vaccine antigen enzyme-linked immunologic detecting kit, described detection kit comprises: pre-coated anti-enterovirns type 71 polyclonal antibody ELISA Plate, sample diluting liquid, second antibody, enzyme labelled antibody, concentrated cleaning solution, zymolyte solution A, zymolyte B solution, stop buffer and antigen standard, ELISA Plate is wrapped by the polyclonal antibody prepared as immunogenic by recombination enterovirus 71 type structural proteins 1 in advance, coating buffer is carbonate buffer solution, package amount is 0.5-2 microgram/hole, bag is closed by the bovine serum albumin(BSA) of rear ELISA Plate 1%-3%, sample diluting liquid is 10 mM/ls of phosphate buffers of bovine serum albumin bletilla 0.1% Tween-20 containing 1%-3%, pH7.2-7.4, second antibody is the monoclonal antibody prepared as immunogenic by keyhole blood blue carrier protein couplet peptide sequence FGEHKQEKDL, doubly uses with sample diluting liquid dilution 500-2000, enzyme labelled antibody is the goat anti-mouse immune globulin antibody of horseradish peroxidase-labeled, doubly uses with sample diluting liquid dilution 500-2000, zymolyte solution A is 3,3', 5,5'-tetramethyl benzidine 0.5g, and acetone 3.5ml, ethanol 46.5ml, aminopyrine 0.2g, citric acid 2.1g, sodium ethylene diamine tetracetate 0.3g, mix after the constant volume to 1000ml that adds water, zymolyte B solution is disodium hydrogen phosphate 36.85g, 30% hydrogen peroxide 155ul, mixes after the constant volume to 1000ml that adds water.Concentrated cleaning solution is for containing 1% Tween-20,100 mM/ls of phosphate buffers of pH7.2-7.4; Stop buffer is the sulfuric acid solution of 2 mol/L; Antigen standard is placed in box.
The preparation method of enterovirns type 71 killed vaccine antigen enzyme-linked immunologic detecting kit, particular flow sheet
See Fig. 1, its concrete steps are as follows:
(1) preparation of anti-enterovirns type 71 polyclonal antibody: with Bacillus coli expression enterovirns type 71 capsid structural protein (VP1) recombinant antigen, purer recombinant antigen is obtained through column chromatography, obtain antigen immune rabbit, gained serum chromatographic column (proteinA) chromatography, is separated the anti-enterovirns type 71 polyclonal antibody after obtaining purifying.
(2) preparation of little mouse-anti enterovirns type 71 monoclonal antibody: with the blue carrier protein of the enterovirns type 71 specific polypeptide FGEHKQEKDL coupling keyhole blood of synthesis as immunogenic immune mouse (product are Balb/c), obtain the mouse boosting cell of antigenic stimulus, the myeloma cell (SP2/0) of this splenocyte and mouse is merged, with the hybridoma positive colony of indirect elisa method screening secreting specificity antibody, then limiting dilution assay subclone carried out to it and identify, obtaining stable monoclonal hybridoma strain.Further the monoclonal hybridoma of in vitro culture is injected in Balb/c mouse peritoneal and prepares ascites, gained ascites proteinA column chromatography, be separated the anti-enterovirns type 71 monoclonal antibody after obtaining purifying.
(3) described enzyme labelled antibody be prepared as commercially available.
(4) preparation of described solution:
Bovine serum albumin bletilla 0.1% Tween-20 of A, sample diluting liquid: 1%-3%, 10 mM/ls of phosphate buffers of pH7.2-7.4.
B, concentrated cleaning solution are for containing 1% Tween-20,100 mM/ls of phosphate buffers of pH7.2-7.4.
C, zymolyte solution A: get 3,3', 5,5'-tetramethyl benzidine 0.5g, acetone 3.5ml, ethanol 46.5ml, aminopyrine 0.2g, citric acid 2.1g, sodium ethylene diamine tetracetate 0.3g, mix after the constant volume to 1000ml that adds water.
D, zymolyte B solution: disodium hydrogen phosphate 36.85g, 30% hydrogen peroxide 155ul, mixes after the constant volume to 1000ml that adds water.
E, stop buffer: get 98% concentrated sulphuric acid 100ml and add in 800ml water and mix and get final product.
(5) described antigen standard is cultivated on Vero cell by enterovirns type 71 vaccine strain, increases, and obtains through freeze thawing, centrifugal extraction and column chromatography.For the demarcation of the antigenic content of enterovirns type 71 inactivated vaccine.
The present invention has the following advantages and effect: resist with the enterovirns type 71 that enterovirns type 71 capsid structural protein (VP1) recombinant antigen of Bacillus coli expression obtains as immunogenic more, enterovirns type 71 inactivated vaccine is detected there is better specificity, greatly can avoid the nonspecific reaction caused by vaccine cultured cell matrix, thus improve the accuracy detected.And using the mouse monoclonal antibody that enterovirns type 71 specific polypeptide FGEHKQEKDL obtains as immunogenic, because of specificity and the stronger immunogenicity thereof in polypeptide site used, make prepared monoclonal antibody have good specificity.The present invention is gone out in mensuration EV71 virus
During live vaccine antigen titre, there is good sensitivity, have linear preferably within the scope of 5.9-750ng/ml, linearly dependent coefficient r
2be greater than 0.99.Kit provided by the present invention is the detection kit that a species specificity is good, quantitatively accurate, highly sensitive, reproducible, measure EV71 viral inactivation vaccine antigen titre simply and easily.
Accompanying drawing explanation
Fig. 1 is present invention process process flow diagram
Fig. 2 is anti-enterovirus type 71 viruses polyclonal antibody electrophoretogram
Fig. 3 is anti-enterovirus type 71 viruses mouse monoclonal antibody electrophoretogram
Fig. 4 is enterovirus type 71 viruses antigen standard electrophoretogram, M:marker; 1. before purifying; 2. after purifying
Fig. 5 is this kit enterovirus type 71 viruses antigen standard examination criteria curve and regression equation.
Embodiment
Following examples are used for explaining and the present invention are described, instead of limit the invention, of the present invention
In the protection domain of spirit and claim, any amendment make the present invention and change, all fall into protection scope of the present invention.
the preparation of the anti-enterovirus type 71 viruses polyclonal antibody of embodiment 1
1) enterovirns type 71 capsid structural protein (VP1) expressing gene is obtained the recombinant expressed bacterium of expressing enterovirns type 71 capsid structural protein by the method for gene clone;
2) express bacterium with gained recombinant bacterium and express enterovirns type 71 capsid structural protein (VP1) recombinant antigen, obtain purer recombinant antigen through column chromatography;
3) gained antigen immune rabbit, interval one week immunity, altogether immunity four times, each immunity 0.5 milligram.Serum is gathered after final immunization surrounding;
4) gained serum proteinA column chromatography, is separated the anti-enterovirns type 71 polyclonal antibody after obtaining purifying.Gained antibody purification is shown in Fig. 2.
the preparation of the anti-enterovirus type 71 viruses mouse monoclonal antibody of embodiment 2
1) the enterovirns type 71 specific polypeptide FGEHKQEKDL synthesized, and carrier protein couplet blue with keyhole blood;
2) gained coupling protein immunity Balb/c mouse, obtains the mouse boosting cell of antigenic stimulus;
3) myeloma cell (SP2/0) of this splenocyte and mouse is merged, with indirect elisa method screening point
Secrete the hybridoma positive colony of specific antibody, then limiting dilution assay subclone carried out to it and identify
IgG hypotype, obtains stable monoclonal hybridoma strain;
4) monoclonal hybridoma of in vitro culture is injected in Balb/c mouse peritoneal and prepares ascites, gained ascites proteinA column chromatography, be separated the anti-enterovirns type 71 monoclonal antibody after obtaining purifying.Gained antibody purification is shown in Fig. 3.
the preparation of embodiment 3 enterovirus type 71 viruses antigen standard
1) Enterovirus 71 antigen standard items are cultivated on Vero cell by enterovirns type 71 vaccine strain, increase, and obtains through freeze thawing, centrifugal extraction and column chromatography.For the demarcation of the antigenic content of enterovirns type 71 inactivated vaccine.Gained antigen is shown in Fig. 4.
the preparation of the pre-coated anti-enterovirns type 71 polyclonal antibody ELISA Plate of embodiment 4
1) with carbonate buffer solution, anti-enterovirns type 71 polyclonal antibody is diluted to 5 ~ 20 mcg/ml, by the amount in 50 ~ 100 microlitres/hole, adds the aerial of ELISA Plate, be put in 4 DEG C of overnight incubation, wash plate 3 ~ 5 times by phosphate buffer washing lotion;
2) add confining liquid with confining liquid by the amount in 200 microlitres/hole, hatch 1 ~ 2 hour for 37 DEG C, phosphate buffer (PBS) washing lotion washes plate 3 ~ 5 times, dries.
Described solution is as follows:
Bag is buffered liquid (0.05M carbonate buffer solution, PH9.6): 1.59 grams, sodium carbonate, sodium bicarbonate 2.93 grams, adding distil water to 1000 milliliter.
Phosphate buffer washing lotion (PBS, PH7.4): potassium dihydrogen phosphate 0.2 gram, sodium hydrogen phosphate 2.9 grams, 8.0 grams, sodium chloride, 0.2 gram, potassium chloride, Tween-200.5 milliliter, adding distil water to 1000 milliliter.
Confining liquid: add 1 gram of bovine serum albumin(BSA) in 100 milliliters of PBS washing lotions, dissolves mixing.
the preparation of other solution of embodiment 5 kit
1) bovine serum albumin bletilla 0.1% Tween-20 of sample diluting liquid: 1%-3%, 10 mM/ls of phosphate buffers of pH7.2-7.4.
2) concentrated cleaning solution is for containing 1% Tween-20,100 mM/ls of phosphate buffers of pH7.2-7.4.
3) zymolyte solution A: get 3,3', 5,5'-tetramethyl benzidine 0.5g, acetone 3.5ml, ethanol 46.5ml, aminopyrine 0.2g, citric acid 2.1g, sodium ethylene diamine tetracetate 0.3g, mix after the constant volume to 1000ml that adds water.
4) zymolyte B solution: disodium hydrogen phosphate 36.85g, 30% hydrogen peroxide 155ul, add water constant volume
Mix after 1000ml.
5) stop buffer: get 98% concentrated sulphuric acid 100ml and add in 800ml water and mix and get final product.
embodiment 6 enterovirns type 71 killed vaccine antigen enzyme-linked immunologic detecting kit
1) first Enterovirus 71 antigen standard items dilution is carried out multiple proportions serial dilution, with micropipettor, the EV71 viral antigen sample diluted and determined antigen sample are added in pre-coated ELISA Plate hole successively, 100 microlitres/hole, each dilute sample adds holes, in the other hole of ELISA Plate, set up negative hole 2 simultaneously, and stay 2 holes to do blank zeroing hole, put into 37 DEG C of water baths and hatch 1 hour, wash plate 4 times by PBS washing lotion;
2) in each hole of above-mentioned ELISA Plate, add the little mouse-anti EV71 viral monoclonal antibodies that dilutability is 1:1000,100 microlitres/hole, puts into 37 DEG C of water baths and hatches 1 hour, wash plate 4 times by PBS washing lotion;
3) in each hole of above-mentioned ELISA Plate, add the enzyme labelled antibody that dilutability is 1:1500,100 microlitres/hole, put into 37 DEG C of water baths and hatch 1 hour, wash plate 5 times by PBS washing lotion;
4) get nitrite ion A liquid 5ml and nitrite ion A liquid 5ml mixes, in each hole of above-mentioned ELISA Plate, add nitrite ion, 100 microlitres/hole, put into wet box, place 20 minutes in room temperature;
5) in each hole of above-mentioned ELISA Plate, add stop buffer, 50 microlitres/hole, be placed in microplate reader under 450nm wavelength, detect absorbance A value with after blank well zeroing.
6) result calculates:
With Enterovirus 71 antigen standard concentration logarithm for y-axis, with corresponding OD450 mean value for x-axis, Criterion curve, calculates regression equation, and typical curve has linear preferably within the scope of 5.9-750ng/ml, linearly dependent coefficient r
2>0.99, the results are shown in Figure 5.
The absorbance (OD450 mean value) of determined antigen sample is substituted in regression equation the content calculating Enterovirus 71 antigen in testing sample.
Claims (4)
1. an enterovirns type 71 killed vaccine antigen enzyme-linked immunologic detecting kit, described detection kit comprises: pre-coated anti-enterovirns type 71 polyclonal antibody ELISA Plate, sample diluting liquid, second antibody, enzyme labelled antibody, concentrated cleaning solution, zymolyte solution A, zymolyte B solution, stop buffer and antigen standard, it is characterized in that ELISA Plate is wrapped by the polyclonal antibody prepared as immunogenic by recombination enterovirus 71 type structural proteins 1 in advance, coating buffer is carbonate buffer solution, package amount is 0.5-2 microgram/hole, bag is closed by the bovine serum albumin(BSA) of rear ELISA Plate 1%-3%, sample diluting liquid is 10 mM/ls of phosphate buffers of bovine serum albumin bletilla 0.1% Tween-20 containing 1%-3%, pH7.2-7.4, second antibody is the monoclonal antibody prepared as immunogenic by keyhole blood blue carrier protein couplet peptide sequence FGEHKQEKDL, doubly uses with sample diluting liquid dilution 500-2000, enzyme labelled antibody is the goat anti-mouse immune globulin antibody of horseradish peroxidase-labeled, doubly uses with sample diluting liquid dilution 500-2000, zymolyte solution A is 3,3', 5,5'-tetramethyl benzidine 0.5g, and acetone 3.5mL, ethanol 46.5mL, aminopyrine 0.2g, citric acid 2.1g, sodium ethylene diamine tetracetate 0.3g, mix after the constant volume to 1000mL that adds water, zymolyte B solution is disodium hydrogen phosphate 36.85g, and 30% hydrogen peroxide 155 μ L, mixes after the constant volume to 1000mL that adds water.
2. enterovirns type 71 killed vaccine antigen enzyme-linked immunologic detecting kit according to claim 1, is characterized in that described concentrated cleaning solution is for containing 1% Tween-20,100 mM/ls of phosphate buffers of pH7.2-7.4.
3. enterovirns type 71 killed vaccine antigen enzyme-linked immunologic detecting kit according to claim 1, is characterized in that described stop buffer is the sulfuric acid solution of 2 mol/L.
4. enterovirns type 71 killed vaccine antigen enzyme-linked immunologic detecting kit according to claim 1, it is characterized in that described antigen standard is cultivated on Vero cell by enterovirns type 71 vaccine strain, increases, obtain through freeze thawing, centrifugal extraction and column chromatography.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410839523.5A CN104569428B (en) | 2014-12-30 | 2014-12-30 | A kind of enterovirns type 71 killed vaccine antigen enzyme-linked immunologic detecting kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410839523.5A CN104569428B (en) | 2014-12-30 | 2014-12-30 | A kind of enterovirns type 71 killed vaccine antigen enzyme-linked immunologic detecting kit |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104569428A CN104569428A (en) | 2015-04-29 |
| CN104569428B true CN104569428B (en) | 2016-03-23 |
Family
ID=53085980
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410839523.5A Active CN104569428B (en) | 2014-12-30 | 2014-12-30 | A kind of enterovirns type 71 killed vaccine antigen enzyme-linked immunologic detecting kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104569428B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105259347B (en) * | 2015-08-13 | 2017-11-14 | 杭州雅盛生物科技有限公司 | The detection method and its kit of a kind of Enterovirus 71 |
| CN105759033A (en) * | 2016-03-29 | 2016-07-13 | 中国医学科学院医学生物学研究所 | Identification detecting method for Cox A16 and EV71 viral antigens |
| CN107037207B (en) * | 2017-06-20 | 2018-08-24 | 山东莱博生物科技有限公司 | A kind of c-hepatitis antibody polymer enzyme marker and its preparation and application |
| CN111458498A (en) * | 2019-01-19 | 2020-07-28 | 艾维可生物科技有限公司 | Hand-foot-mouth EV71 antigen detection kit |
| CN110078821B (en) * | 2019-03-20 | 2022-03-25 | 天津大学 | Sequence of enterovirus D group 68 type VP1 monoclonal antibody and application thereof |
| CN117209571A (en) * | 2023-08-23 | 2023-12-12 | 北京清华长庚医院 | VP1 antigen, kit and method for detecting BKV virus VP1 antibody |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI487537B (en) * | 2010-06-09 | 2015-06-11 | Nat Health Research Institutes | Monoclonal antibodies against enterovirus 71 and uses thereof |
| CN101881774A (en) * | 2010-06-22 | 2010-11-10 | 浙江普康生物技术股份有限公司 | Detection method for measuring titer of antigens of enterovirus71 inactivated vaccine |
| CN102539776A (en) * | 2010-12-10 | 2012-07-04 | 浙江普康生物技术股份有限公司 | Enzyme-linked immunosorbent assay method for enterovirus 71-specific antibody |
| CN104220090A (en) * | 2011-09-20 | 2014-12-17 | 淡马锡生命科学实验室有限公司 | Enterovirus 71 specific antibodies and uses thereof |
| CN102854317A (en) * | 2011-09-27 | 2013-01-02 | 上海博沃生物科技有限公司 | EV71 (human enterovirus 71) antigen enzyme-linked reaction detection kit and its preparation method |
| CN102854312A (en) * | 2012-07-31 | 2013-01-02 | 上海博沃生物科技有限公司 | Enzyme-linked reaction detection kit for EV71 antigens |
-
2014
- 2014-12-30 CN CN201410839523.5A patent/CN104569428B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN104569428A (en) | 2015-04-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104569428B (en) | A kind of enterovirns type 71 killed vaccine antigen enzyme-linked immunologic detecting kit | |
| Westdijk et al. | Characterization and standardization of Sabin based inactivated polio vaccine: proposal for a new antigen unit for inactivated polio vaccines | |
| CN103076451B (en) | A kind of O type aftosa 146S antigen quantify ELISA detection kit and using method thereof | |
| CN102305859B (en) | Activated toxic antibody kit for detecting porcine reproductive and respiratory syndrome virus | |
| CN105675866B (en) | For detecting that the solid phase of O-shaped FMDV antibody blocks ELISA kit | |
| CN111072776A (en) | Monoclonal antibody against SpA5 protein, application thereof and kit containing monoclonal antibody | |
| CN101936997B (en) | Human anti-rabies virus IgG antibody ELISA test kit | |
| CN113265006B (en) | Fusion protein 3AN for capturing PRRSV nucleocapsid protein antibody and application thereof | |
| WO2012163263A1 (en) | Reagents and methods for prrsv detection | |
| Wang et al. | A neutralizing monoclonal antibody-based competitive ELISA for classical swine fever C-strain post–vaccination monitoring | |
| CN105067818A (en) | ELISA (enzyme linked immunosorbent assay) antibody detection kit for O type foot-and-mouth disease virus and detection method adopting ELISA antibody detection kit | |
| CN113336845A (en) | Porcine single B cell antibody of PRRSV nucleocapsid protein and competitive ELISA antibody detection kit | |
| CN106771181A (en) | A kind of bird flu H7N9 hemagglutinin HA antigens detect ELISA kits and detection method | |
| Ran et al. | Development and validation of a competitive ELISA based on bacterium-original virus-like particles of serotype O foot-and-mouth disease virus for detecting serum antibodies | |
| CN101082626A (en) | ELISA reagent kit for testing Asial type foot-and-mouth disease virus | |
| CN105004856A (en) | Porcine circovirus II antibody detection ELISA kit | |
| CN102854317A (en) | EV71 (human enterovirus 71) antigen enzyme-linked reaction detection kit and its preparation method | |
| CN111537711B (en) | Kit for rapidly and accurately detecting adenovirus type 3 and preparation method thereof | |
| Njayou et al. | Purification of measles virus by affinity chromatography and by ultracentrifugation: a comparative study | |
| CN109374886B (en) | Infectious bovine rhinotracheitis virus antibody detection kit and application thereof | |
| CN103293311A (en) | Indirectly competitive ELISA (Enzyme Linked Immunosorbent Assay) immune kit for detecting porcine reproductive and respiratory syndrome virus | |
| Onodera et al. | Immune-focusing properties of virus-like particles improve protective IgA responses | |
| CN101881774A (en) | Detection method for measuring titer of antigens of enterovirus71 inactivated vaccine | |
| CN109613249A (en) | A kind of russian spring-summer encephalitis virus IgM antibody ELISA detection kit and preparation method thereof | |
| CN104744590A (en) | A monoclonal antibody for H1N1 swine influenza A virus hemagglutinin protein and a double-antibody sandwich ELISA kit |
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 |