CN101362800A - Test strip for rapid detection of brucella - Google Patents
Test strip for rapid detection of brucella Download PDFInfo
- Publication number
- CN101362800A CN101362800A CNA2008101127305A CN200810112730A CN101362800A CN 101362800 A CN101362800 A CN 101362800A CN A2008101127305 A CNA2008101127305 A CN A2008101127305A CN 200810112730 A CN200810112730 A CN 200810112730A CN 101362800 A CN101362800 A CN 101362800A
- Authority
- CN
- China
- Prior art keywords
- monoclonal antibody
- test strip
- brucella
- pad
- antibody
- 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
Images
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention provides a quick test dipstick used for testing a specific antigen of Brucella, which comprises a reaction membrane and a binder release liner. The reaction membrane is provided with a testing strap coated with a bp26 monoclonal antibody or bp26 polyclonal antibody and a quality control strap coated with anti-IgG; the binder release liner is coated with the bp26 monoclonal antibody which is marked by colloidal gold to be different from an antigen binding site on the reaction membrane. A membrane chromatography double antibody sandwich method is applied for detecting the specific antigen of the Brucella in a specimen. As the dipstick is adopted for test, the operation is simple, convenient, quick and concise, requiring no special equipment and facilities as well as professional training. Furthermore, the dipstick has clear and easy-identity results, simple operation and easy popularization, is applicable to matrixes, field tests of emergency on a large scale and the study of epidemiology and can aid in the infection diagnostics of the Brucella.
Description
Technical field
The invention belongs to field of biological detection, relate to the preparation and the Study of Monoclonal Antibodies thereof of brucella outer membrane protein bp26 gene engineering antigen and contain the brucella colloidal gold fast detecting test paper strip and the application thereof of this monoclonal antibody.
Technical background
Brucellosis (brucellosis) is the beastly transmissible disease of suffering from altogether of people that is caused by Brucella (Brucella), is commonly called as brucellosis.Clinical characters is long-term heating, hidrosis, arthralgia, tired, hepatosplenomegaly etc., and this disease all has in various degree popular in countries in the world.
The people is mainly by skin, mucous membrane, digestive tube and respiratory tract infection, especially with infect brucella melitensis, ox kind brucella is the most serious.Pig kind brucella infected person is more rare, and kind of dog brucella infected person is rare, and sheep epididymis kind brucella, sarin mouse kind brucella be infected person not substantially.
The easy mistaken diagnosis of brucellosis is the long-term heating cause diagnosis and treatment to be looked into of rheumatism, typhoid fever, tuberculosis, virus infection or conduct.The major cause of its long-term misdiagnosis is: (1) clinician is the major cause that causes mistaken diagnosis to the understanding deficiency of brucellosis.(2) the epidemiologic data inquiry is not detailed, particularly medical history, contact history, occupation, food habits, residence and popular area etc.(3) clinical manifestation variation and atypical symptoms.(4) lack simple to operate, quick, special, responsive detection means.
At present the brucella INFECTION IN DETECTION is mainly carried out in the laboratory.Mainly contain:
(1) etiological diagnosis:
Microscopy: gather tissues such as miscarriage afterbirth, chorion edematous fluid, liver, spleen, lymphoglandula, fetus gastric content, make and smear sheet, with the dyeing of Ke Ziluo Paderewski staining, microscopy, brucella is red club shape dialister bacterium, and other bacterium is blue.
Separation and Culture: fresh pathological material of disease can be with culture medium culturing such as tryptose agar face or blood agar inclined-plane, liver soup agar slant, 3% glycerine, 0.5% glucose liver soup agar slants; Cultivate after 7～10 days, carry out colony characteristics inspection and monospecific antiserum agglutination test for 37 ℃.
(2) serodiagnosis: main following several, brave red plate agglutination test (RBPT) full milk ring test (MRT), tube agglutination test (SAT), complement fixation test (CFT) (CFT) etc.In recent years, people detect the antibody that infects in the serum with the brucellar antigen development ELISA method detection kit of slightly carrying, and have obtained good effect.
(3) other detects the specific specificity Nucleotide of brucella etc. as PCR.
Bp26 albumen is a kind of of brucella outer membrane protein, existing studies show that, bp26 albumen has good immunogenicity, and from gene level, various brucella (B.abortus, B.ovis and B.melitensis etc.) the bp26 gene order almost completely consistent, just Nucleotide has fine difference, but the aminoacid sequence indifference.Therefore belong to various brucellar common antigens, can be used as brucellar detection antigen, be used for detecting diagnosis.
Summary of the invention
The objective of the invention is to utilize various brucellar common outer membrane protein bp26, develop specific monoclonal antibody; And utilize this antigenic monoclonal antibody, develop a kind of can be quick, accurately check brucellar quick detection test paper (colloidal gold method).
Another object of the present invention is to provide the preparation method of above-mentioned test strip.
For achieving the above object, the present invention at first utilizes genetic engineering technique, clones and efficiently express brucella outer membrane protein antigen bp26, and by immunological technique its immunogenicity and specificity is examined and determine.Then, utilize bp26 to develop the specific pairing monoclonal antibody of brucella.Experiment shows that by the monoclonal antibody of gene engineering antigen preparation, it has good specificity and sensitivity.
And then, the invention provides a kind of test strip, comprise that reaction film and binding substances discharge pad, described reaction film have bag by the detection band of bp26 monoclonal antibody or bp26 polyclonal antibody and bag by the quality control band of two anti-IgG; Described binding substances discharge the pad bag by colloid gold label with reaction film on the bp26 monoclonal antibody of different antigen binding sites.
Wherein, reaction film can be nitrocellulose membrane, and binding substances discharges pad and can be glass fibre membrane.
Wherein, two anti-IgG are anti-mouse IgG antibody.
The present invention also provides a kind of method for preparing above-mentioned test strip, and it comprises the steps:
1) prepare monoclonal antibody according to the method described above, or the preparation polyclonal antibody;
2) monoclonal antibody that step 1) is prepared or polyclonal antibody and two anti-IgG form respectively on reaction film and detect band and quality control band, and are standby;
3) with the monoclonal antibody of colloid gold label claim 1 preparation, bag is discharged in the pad to binding substances, wherein the monoclonal antibody and step 2 of colloid gold label) in to be coated on the antigen binding site that detects the monoclonal antibody in being with different;
4) with the reaction film for preparing and binding substances discharges pad and sample pad, absorbent pad and reaction upholder are assembled into test strip.
Brucellergen quick detection test paper of the present invention (colloidal gold method) has the following advantages:
(1) detects fast: went out the result in 10 minutes;
(2) specificity: only brucella is positive, and to other germs such as salmonella typhi, tubercule bacillus, etc. the pathogenic agent result that is negative;
(3) susceptibility: bp26 detects the level that reaches 1ng to the brucella outer membrane protein;
(4) storage and transport are convenient, at room temperature can preserve 18 months;
(5) be convenient to clinical and the household use, and have industry.
Description of drawings
Fig. 1: the front schematic view of A test strip of the present invention; The side schematic view of B test strip of the present invention.Wherein, 1: absorbent pad; 2: nitrocellulose membrane (T: the band of brucellar monoclonal antibody or anti-brucellar polyclonal antibody; C: bag is by the Quality Control band of anti-mouse IgG); 3: the glass fibre membrane that contains the brucellar monoclonal antibody of colloid gold label; 4: golden labeling antibody protective membrane; 5: the reaction upholder.
Fig. 2: detected result synoptic diagram.Wherein, be followed successively by from left to right: two line positives of T, C; Line feminine gender of C; Two line feminine genders of T, C are invalid.
Embodiment
Following examples further specify content of the present invention, but should not be construed as limitation of the present invention.Without departing from the spirit and substance of the case in the present invention, modification or replacement to the inventive method, step or condition are done all belong to scope of the present invention.
If do not specialize the conventional means that used technique means is well known to those skilled in the art among the embodiment.
Embodiment 1: the preparation of brucella outer membrane protein bp26 gene engineering antigen
(1) acquisition of goal gene
The primer that contains the restricted interior enzyme EcoR1 of association, Xho1 restriction enzyme site according to the characteristics design two ends of target gene fragment sequence (the GenBank accession number is AY166769) and pGEX-4T-1 (Pharmacia) expression vector:
5’gaattcatgaacactcgtgct3’
5’gcctcgagttacttgatttcaa3’
Then, from the brucella genome, amplify goal gene segment bp26, amplification condition: 95 ℃ of sex change 5min; 95 ℃ of 1min, 49.8 ℃ of 1min, 70 ℃ of 1min carry out 35 circulations; Last 70 ℃ are extended 10min.
(2) screening of the clone of goal gene and positive recombinant
Cut glue behind the pcr amplification product electrophoresis and reclaim, with the PMD-18T cloning vector spend the night for 16 ℃ be connected after, be transformed in the DH5 α competent cell, picking mono-clonal bacterial strain, 37 ℃ of incubated overnight are behind the extraction plasmid, with the plasmid is that template is carried out PCR evaluation positive colony bacterial strain, measures sequence.
(3) structure of bp26 fusion expression vector
With restriction enzyme EcoRI, XhoI respectively enzyme cut T/bp26 and pGEX-4T-1,1% agarose electrophoresis is cut the big fragment after glue reclaims bp26 purpose fragment and pGEX-4T-1 double digestion, with the T4 ligase enzyme with both 16 ℃ of connections of spending the night, connect product and change the BL21 competent cell over to, the LB solid medium was cultivated 8～10 hours, extract plasmid after the picking list bacterium colony overnight incubation, identify respectively with PCR and restriction enzyme EcoR I, XhoI double digestion, PCR product and enzyme are cut product and are analyzed with 1% agarose electrophoresis, screening positive clone is with the plasmid order-checking of reorganization.
(4) abduction delivering of pGEX-bp26 fusion rotein
The positive colony bacterium that filters out shakes the bacterium overnight incubation in the LB substratum after, the bacterium that will spend the night is seeded in the 1000ml LB liquid nutrient medium according to the ratio of 1:100, and 37 ℃ of shaking tables are cultivated.Bp26 gene transformation bacterium is induced opportunity (OD600nm 0.5) back 2 hours of inoculation for the best, and dense ℃ of 0.3mmol/L of IPTG induced 8 hours for 29 ℃, and target protein is present in the cell pyrolysis liquid supernatant.
(5) purifying of pGEX-bp26 fusion rotein, evaluation
1000ml bacterium liquid 12000r/m in (4), 4 ℃ is centrifugal, abandon supernatant, with thalline cell pyrolysis liquid cracking, use contains the affinity chromatography pillar purified fusion protein of GST label, (B-PER bacterium GST tag fusion protein pillar purifying Kit article No.: the silent generation that science and technology that flies of 78200 matches), carry out purifying by test kit recommendation step and system, purified product carries out the SDS-PAGE electrophoresis to judge purification effect, records productive rate more than 95% by the ultraviolet thin layer.
Western-blot identifies the immunocompetence of fusion rotein.Cutting half glue changes on nitrocellulose filter with the half-dried electroporation that BioRad company produces, and carries out the trace test, and one anti-ly is Leptospira monoclonal antibody (Beijing Bo Aosen Bioisystech Co., Ltd), and after the TMB colour developing, the result has the specific proteins band to occur.
Embodiment 2: anti-brucella outer membrane protein bp26 MONOCLONAL ANTIBODIES SPECIFIC FOR
(1) immune mouse
After the gene engineering antigen of preparation takes out dissolving from-20 cryogenic refrigerators, give BALB/C mice back subcutaneous injection (0.2ml/ only), 10 days at interval.Merged preceding 3 days, and attacked with antigen 0.15ml at mouse peritoneal.Immune effect detects with the ELISA method.
(2) myeloma cell
SP2/0 myeloma cell: purchase in Microbiology and Epidemic Disease Inst., Academy of Military-Medical Sciences (C.Time spent will be stored in the SP2/0 cell recovery in the liquid nitrogen container, cultivate in containing 10% calf serum DMEM substratum 48-72 hour, treat the cell well-grown, perfectly round, bright, the big or small homogeneous of cell, marshalling, be the logarithm division, prepare to merge.
(3) cytogamy
The splenic lymphocyte of the BALB/C mice of SP2/0 cell for preparing and immunity is mixed with 2 * 10 respectively
7/ ml and 1 * 10
8/ ml.Respectively get under the 1ml room temperature and mix.Use 0.8ml, 50% PEG (molecular weight 1500) is as fusogen; Nutrient solution is with 10% calf serum DMEM.
(4) detection in positive hole and screening
The ELISA method detects.Bag is by the rough antigen of brucella and two kinds of enzyme plates of bp26 proteantigen.Culture supernatant to fused cell detects, and selects the ELISA OD value of tiring to carry out subclone greater than 1.0 positive hole.Detect positive colony 27 strains altogether, select 9 strains to carry out the subclone operation.
(5) subclone of monoclonal antibody cell strain: the positive colony cell strain is carried out the cloning screening with limiting dilution assay.To screening positive colony through frozen, process such as recover, go down to posterity, but finally obtain the cell strain of monoclonal antibody of the anti-brucella outer membrane protein of 6 strain stably excreting specificitys bp26.
(6) cell strain of monoclonal antibody calibrating
The cell strain karyotyping: it is 96 that above-mentioned hybridoma karyomit(e) is detected; Prove that they are hybridomas of SP2/0 myelomatosis and mouse cell.
Cell strain stability: the hybridoma cell strain that 6 strains is produced brucella outer membrane protein bp26 monoclonal antibody detects monoclonal antibody positive rate 100% through continuous cloning, subculture in vitro separately 5 months, and all can reach the secretory antibody that keeps stable through cryopreservation resuscitation repeatedly, culture supernatant 1:400-1:500 (++) IFA that tires.
Embodiment 3: the calibrating of anti-Brucella outer membrane protein bp26 monoclonal antibody
(1) odd contradictive hydroperitoneum preparation:
Mouse: SPF level mouse, do not have mouse source virus pollution on inspection, in the ascites production process as find that animal is unhealthy, bite, the infected should discard.
The cell strain enlarged culturing: get and produce batch 1 recovery of cell pipe, Ensure Liquid liquid enlarged culturing is produced cell for 1 and only used once, and is no longer frozen.
The hybridoma cell strain inoculation: preparation ascites all needs to carry out under aseptic condition, before the injection hybridoma, and every mouse peritoneal injecting fluid paraffin 0.5ml.One every the mouse peritoneal injection in week back hybridoma 1-3 * 10
6/ 0.2ml.
Ascites is gathered: injection cell strain 7-10 days, or mouse once gathers ascites before dying, puts-20 ℃ of preservations.
(2) purification of monoclonal antibody:
Adopt ammonium sulfate precipitation method slightly to carry, and then,, only show single protein band through the calibrating of SDS-PAGE electrophoresis with HiTrap rProtein A post (GE HealthcareLife Sciences) purifying.
(3) antibody affinity is measured: adopt the experiment of NaSCN competitive ELISA, measure the drop-out value of its ED50, reflect the size of its antibody affinity indirectly.Select wherein avidity 6 strains preferably, carry out the hypotype calibrating.
(4) hypotype calibrating: adopt the monoclonal antibody of immunodiffusion method pair cell culture supernatant to carry out the detection of Ig subclass.The Ig subclass of above-mentioned 6 strain monoclonal antibodies is respectively: IgG1 has three strains; IgG2a has two strains; IgG2b has a strain.
(5) antigen site calibrating: detect with the antigen binding site of addition ELISA method to 6 strain monoclonal antibodies.The result shows that four kinds of different antigen binding sites are arranged in this 6 strain.
Embodiment 4: the establishment of brucellergen test strip
(1) preparation of Radioactive colloidal gold-antibody conjugates:
Definite through testing, the bp26 monoclonal antibody that makes with embodiment 3 is as golden labeling antibody, and its best combination pH value is 8.0, and the albumen proportioning is 29 μ g/ml Radioactive colloidal golds.The mark Radioactive colloidal gold by the amount of every square centimeter 65 μ l, is got Radioactive colloidal gold-antibody conjugates solution after stablizer (0.5%BSA, pH8.0,0.01M Tris damping fluid) is handled, evenly is adsorbed on the glass fibre, and lyophilize, and in dry environment, preserve.
(2) coated antibody is in nitrocellulose membrane:
With brucella outer membrane protein bp26 monoclonal antibody (with colloid gold label when the clonal antibody antigen binding site different) be diluted to 1.5 ± 0.1mg/ml with 0.01M PBS.The sheep anti-mouse igg polyclonal antibody is diluted to 2 ± 0.1mg/ml with 0.01M PBS.With Membrane jetter the two speed with 1 μ l/cm is sprayed on the nitrocellulose membrane, forms detection line and control line respectively.Article two, be spaced apart 0.5cm between the line.
The cellulose nitrate that is fixed with antibody was put in 37 ℃ of baking boxs dry 2 hours.Preserve standby in the dry environment.
(3) the brucellergen detection kit is formed
The reaction upholder is 6.5cm * 0.4cm PCV plate; Absorbent pad is the filter paper for oil of 2cm * 0.4cm; 1.8cm the nitrocellulose membrane of * 0.4cm wraps successively by anti-mouse IgG, proteic monoclonal antibody of brucella bp26 (step 2) or anti-brucellar polyclonal antibody, the proteic monoclonal antibody glass fibre of the brucella bp26 (step 1) that contains colloid gold label of 0.4cm * 0.4cm; Gold labeling antibody protective membrane (sample pad) is the filter paper fibre of 2.7cm * 0.4cm; Promptly formed brucellergen quick detection test paper bar (colloidal gold method).
(4) brucellergen detection kit specificity and susceptibility experiment:
Use the detection kit (detect band and be monoclonal antibody) of step 3 to experimentize
The specificity experiment: the following negative quality control product of brucellergen detection kit specificity checking design comprises intestinal bacteria, dysentery bacterium, campylobacter jejuni, Pestivirus suis, pork measles antigen, Salmonella typhimurium, Vibrio parahaemolyticus, Leptospira.Detected result shows that this product no cross reaction equal with it has good specificity.
The susceptibility experiment: detect ox kind cloth Salmonella, pig kind cloth Salmonella, sheep kind cloth Salmonella, the kind of dog cloth Salmonella of different concns, its limit of identification is 1 * 10
6-1 * 10
7Bacterium/ml detects bp26 albumen, can reach 5ng/ml.
Sample 100-150 μ l to be checked is added on the sample pad (embodiment 4 test strip " 4 " are located) of detector bar, sample liquid is upwards creeped along film, 10-15 minute sentence read result.
The result:
As containing brucellergen in the sample, then with test strip on the brucellar monoclonal antibody of colloid gold label form corresponding mixture, up and the brucellar monoclonal antibody or the anti-brucellar polyclonal antibody that are coated on the nitrocellulose membrane, form red lines, promptly form red stripes, be positive findings at the T place.
No matter whether contain corresponding antigen, the brucellar monoclonal antibody of colloid gold label continues upwards to creep and the anti-mouse IgG that is coated on the film forms the red precipitate line, promptly locates to form red stripes at " C ".This line is a nature controlling line, loses efficacy as Radioactive colloidal gold, and this line just can not occur, and illustrates that test strip lost efficacy.
Sequence table
＜110〉Beijing Zhuangdi Haohe Biomedicine Science and Technology Co., Ltd
＜120〉a kind of test strip that is used for the brucella rapid detection
<130>
<160>2
<170>PatentIn?version?3.3
<210>1
<211>21
<212>DNA
＜213〉artificial sequence
<400>1
<210>2
<211>22
<212>DNA
＜213〉artificial sequence
<400>2
Claims (10)
1, anti-brucella outer membrane protein bp26 monoclonal antibody is characterized in that, being prepared by the bp26 gene engineering antigen of described monoclonal antibody.
2, the described monoclonal antibody method of a kind of preparation claim 1, it comprises step: with construction expression bp26 expression carrier, with described expression vector transformed host cell, abduction delivering bp26 albumen, purified back is as the immunogen preparing monoclonal antibody.
3, method as claimed in claim 2, it also comprises the antigen binding site of the monoclonal antibody that detects preparation.
4, the detection kit that contains the described monoclonal antibody of claim 1.
5, a kind of test strip comprises that reaction film and binding substances discharge pad, it is characterized in that, described reaction film have bag by the detection band of described monoclonal antibody of claim 1 or bp26 polyclonal antibody and bag by the quality control band of two anti-IgG; Described binding substances discharge the pad bag by colloid gold label with reaction film on the bp26 monoclonal antibody of different antigen binding sites.
6, test strip as claimed in claim 5 is characterized in that, described reaction film is a nitrocellulose filter.
7, test strip as claimed in claim 5 is characterized in that, described binding substances discharges pad and is glass fibre membrane.
8, as claim 5～7 test strip as described in each, it is characterized in that described two anti-IgG are anti-mouse IgG.
9, a kind of method for preparing the arbitrary described test strip of claim 5～8 comprises step:
1) prepare monoclonal antibody according to claim 2 or 3 described methods, or the preparation polyclonal antibody;
2) monoclonal antibody that step 1) is prepared or polyclonal antibody and two anti-IgG form respectively on reaction film and detect band and quality control band, and are standby;
3) with the monoclonal antibody of colloid gold label claim 1 preparation, bag is discharged in the pad to binding substances, wherein the monoclonal antibody and step 2 of colloid gold label) in to be coated on the antigen binding site that detects the monoclonal antibody in being with different;
4) with the reaction film for preparing and binding substances discharges pad and sample pad, absorbent pad and reaction upholder are assembled into test strip.
10, the application of each described test strip of claim 5～8 in detecting brucellergen.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008101127305A CN101362800A (en) | 2008-05-26 | 2008-05-26 | Test strip for rapid detection of brucella |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008101127305A CN101362800A (en) | 2008-05-26 | 2008-05-26 | Test strip for rapid detection of brucella |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101362800A true CN101362800A (en) | 2009-02-11 |
Family
ID=40389423
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2008101127305A Pending CN101362800A (en) | 2008-05-26 | 2008-05-26 | Test strip for rapid detection of brucella |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101362800A (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102432673A (en) * | 2011-12-14 | 2012-05-02 | 南方医科大学 | Brucella bp26 protein epitope, monoclonal antibody and application thereof |
| CN102584957A (en) * | 2012-02-27 | 2012-07-18 | 江苏出入境检验检疫局动植物与食品检测中心 | Specific antibody of Brucella specificity multi-epitope artificial polypeptide, immunomagnetic beads coated with specific bodies, and application of beads |
| CN103149356A (en) * | 2013-01-29 | 2013-06-12 | 杭州迪恩科技有限公司 | Test paper card for testing Brucella antibody through sandwich method |
| CN104749362A (en) * | 2015-03-09 | 2015-07-01 | 中国农业科学院特产研究所 | Deer brucellosis colloidal gold antibody detection test paper strip and preparation method thereof |
| CN106749562A (en) * | 2016-11-30 | 2017-05-31 | 中国兽医药品监察所 | A kind of gene expression product BLSJ 1 of brucella diagnostic marker effect and preparation method thereof |
| CN106749563A (en) * | 2016-11-30 | 2017-05-31 | 中国兽医药品监察所 | A kind of gene expression product BLSJ 3 of brucella diagnostic marker effect and preparation method thereof |
| CN107286250A (en) * | 2017-06-14 | 2017-10-24 | 杭州亿米诺生物科技有限公司 | A kind of brucella fusion protein, its preparation method and application |
| CN109655614A (en) * | 2019-02-26 | 2019-04-19 | 广州维佰生物科技有限公司 | Ox brucellosis virus fluorescence micro-ball immune chromatography test paper and preparation method thereof and detection method |
| CN111879925A (en) * | 2019-12-30 | 2020-11-03 | 杭州奥泰生物技术股份有限公司 | Colloidal gold immunochromatographic assay test paper for rapidly diagnosing brucellosis |
-
2008
- 2008-05-26 CN CNA2008101127305A patent/CN101362800A/en active Pending
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102432673A (en) * | 2011-12-14 | 2012-05-02 | 南方医科大学 | Brucella bp26 protein epitope, monoclonal antibody and application thereof |
| CN102432673B (en) * | 2011-12-14 | 2013-07-03 | 南方医科大学 | Brucella bp26 protein epitope, monoclonal antibody and application thereof |
| CN102584957B (en) * | 2012-02-27 | 2014-02-19 | 江苏出入境检验检疫局动植物与食品检测中心 | Specific antibody of Brucella specificity multi-epitope artificial polypeptide, immunomagnetic beads coated with specific bodies, and application of beads |
| CN102584957A (en) * | 2012-02-27 | 2012-07-18 | 江苏出入境检验检疫局动植物与食品检测中心 | Specific antibody of Brucella specificity multi-epitope artificial polypeptide, immunomagnetic beads coated with specific bodies, and application of beads |
| CN103149356B (en) * | 2013-01-29 | 2016-03-16 | 浙江迪恩生物科技股份有限公司 | A kind of Test paper card utilizing sandwich method to detect Brucella abortus antigen |
| CN103149356A (en) * | 2013-01-29 | 2013-06-12 | 杭州迪恩科技有限公司 | Test paper card for testing Brucella antibody through sandwich method |
| CN104749362A (en) * | 2015-03-09 | 2015-07-01 | 中国农业科学院特产研究所 | Deer brucellosis colloidal gold antibody detection test paper strip and preparation method thereof |
| CN106749562A (en) * | 2016-11-30 | 2017-05-31 | 中国兽医药品监察所 | A kind of gene expression product BLSJ 1 of brucella diagnostic marker effect and preparation method thereof |
| CN106749563A (en) * | 2016-11-30 | 2017-05-31 | 中国兽医药品监察所 | A kind of gene expression product BLSJ 3 of brucella diagnostic marker effect and preparation method thereof |
| CN106749563B (en) * | 2016-11-30 | 2020-05-19 | 中国兽医药品监察所 | Gene expression product BLSJ-3 with brucella diagnosis and identification functions and preparation method thereof |
| CN106749562B (en) * | 2016-11-30 | 2020-06-09 | 中国兽医药品监察所 | Gene expression product BLSJ-1 with brucella diagnosis and identification functions and preparation method thereof |
| CN107286250A (en) * | 2017-06-14 | 2017-10-24 | 杭州亿米诺生物科技有限公司 | A kind of brucella fusion protein, its preparation method and application |
| CN109655614A (en) * | 2019-02-26 | 2019-04-19 | 广州维佰生物科技有限公司 | Ox brucellosis virus fluorescence micro-ball immune chromatography test paper and preparation method thereof and detection method |
| CN111879925A (en) * | 2019-12-30 | 2020-11-03 | 杭州奥泰生物技术股份有限公司 | Colloidal gold immunochromatographic assay test paper for rapidly diagnosing brucellosis |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101362800A (en) | Test strip for rapid detection of brucella | |
| CN109254155A (en) | A kind of detection African swine fever virus antigen colloidal gold immune chromatography test paper and preparation method and application | |
| CN109709339B (en) | Colloidal gold immunochromatographic test strip for detecting skeletal muscle troponin I of cattle or sheep and application thereof | |
| CN101363859B (en) | Test paper strip for rapidly detecting brucellosis antibody | |
| CN107045062B (en) | Detect colloidal gold immuno-chromatography test paper strip, the kit and preparation method thereof of human neutrophil genatinase associated lipocalin | |
| CN101363867B (en) | Test paper strip for detecting clostridium difficile toxin A and toxin B colloidal gold, method for making same | |
| CN105061602B (en) | For detecting fusion protein, the preparation method and application of anti-pig enterotoxigenic escherichia coil antibody | |
| CN101362801A (en) | Rapid detection test strip for detecting campylobacter jejuni specific antigen | |
| CN109705216B (en) | Monoclonal antibody for resisting bovine skeletal muscle troponin I and application thereof | |
| CN109810191B (en) | Monoclonal antibody for resisting sheep skeletal muscle troponin I and application thereof | |
| CN109239341B (en) | Indirect ELISA kit for detecting bovine haemolytic mannheimia antibody and application thereof | |
| CN106636004A (en) | TMV-CMV-PVY three-virus (tobacco mosaic virus, cucumber mosaic virus and potato virus) colloidal gold rapid test strip | |
| CN104062430B (en) | A kind of kit for detecting influenza virus in sample and detection method thereof and application | |
| CN103848916B (en) | Preparation method, coded sequence and the application thereof of a kind of anti-CP4 EPSPS monoclonal antibody | |
| CN111505289A (en) | Peste des petits ruminants detection kit | |
| CN101363862A (en) | Test paper strip for rapidly detecting brucellosis IgM antibody colloidal gold | |
| CN111560341A (en) | Generic inert vector escherichia coli and potential application thereof | |
| CN101363865B (en) | Test paper strip for detecting PRRSV antibody colloidal gold, method for making same and application | |
| CN103823062B (en) | Salmonella choleraesuls enzyme-linked immunologic detecting kit | |
| CN111220809B (en) | Colloidal gold immunochromatography test strip for detecting chicken or duck skeletal muscle troponin I and preparation method and application thereof | |
| CN105223362B (en) | Campylobacter jejuni enzyme-linked immunologic detecting kit | |
| CN106479984A (en) | A kind of dual virus colloidal gold Rapid detection test strip of CMV PVY | |
| CN106198974A (en) | The ELISA detection kit of IHNV antibody in a kind of quick detection rainbow trout serum | |
| CN101363863A (en) | Test paper strip for detecting swine chain coccus type 2 antibody colloidal gold, method for making same and applications | |
| CN106841606B (en) | Detect colloidal gold immuno-chromatography test paper strip, the kit and preparation method thereof of PCT |
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 |
Open date: 20090211 |