CN102879584A - Human blood HSP70 antibody colloidal gold-labeled detection test strip and preparation method thereof - Google Patents
Human blood HSP70 antibody colloidal gold-labeled detection test strip and preparation method thereof Download PDFInfo
- Publication number
- CN102879584A CN102879584A CN201210365239XA CN201210365239A CN102879584A CN 102879584 A CN102879584 A CN 102879584A CN 201210365239X A CN201210365239X A CN 201210365239XA CN 201210365239 A CN201210365239 A CN 201210365239A CN 102879584 A CN102879584 A CN 102879584A
- Authority
- CN
- China
- Prior art keywords
- antigen
- hsp70
- antibody
- colloidal gold
- gold mark
- 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
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a human blood HSP70 antibody colloidal gold-labeled detection test strip and a preparation method thereof, which belongs to the technical field of immunodetection analysis; the test strip comprises a sample pad, a colloidal gold-labeled pad, a cellulose nitrate membrane and absorbent paper orderly connected on a fixed plate; three detection line antigens and one quality control line antibody are disposed on the cellulose nitrate membrane; a colloidal gold-labeled antigen is disposed on the colloidal gold-labeled pad. The colloidal gold-labeled antigen is a developing antigen; the detection line antigen is a capture antigen; the developing antigen and the capture antigen can simultaneously bond with the human HSP70 antibody to form a double antigen sandwich; the developing antigen and the capture antigen are human HSP70 protein obtained by prokaryotic recombinant expression. The test strip of the invention is applicable to human blood HSP70 antibody detection, and has high specificity; the result is easily observed and determined, can be determined within 20 min; the test strip has good stability and repeatability; the operation is simple; no special instrument or equipment is necessary; and the test strip is suitable for popularization and application in on-site detection primary hospitals and experiment research.
Description
Technical field
The invention belongs to the immune detection analysis technical field, be specifically related to HSP70 antibody colloidal gold mark detection test paper bar and preparation method in a kind of human body fluid.
Background technology
Heat-shock protein family (HSPs) is the albumen of a class high conservative, HSP70(heat shock protein 70, heat shock protein) be the important member in the heat-shock protein family, it is divided into the composing type of induction type.When body was subject to that multiple undesirable element stimulates in the external environment, the HSP70 protein expression of induction type raise.HSP70 albumen to the reparation of cellular damage, to survive and keep normal cell function be essential.Cause performance molecular chaperones effect in the cellular stress damage in environmental stimulus, pathogenic bacterial infection, disease, stop protein aggregation, promote the again folding of injury protein.Most mammal, only induction type HSP70 expresses under stressed condition, and just can detect behind significant cell and the physical stress, but people and primate, the HSP70 of induction type has a reference value, the infection that under some specific conditions, causes such as overheated, chemical noxious material exposure, anoxic, nullisomic, infection, autoimmune disease, apoptosis, organ transplant, bacterium and virus, in the heart body pipe diseases such as atherosclerotic, the HSP70 of biosome expresses significantly and raises; In the normal aging process, spermatogenesis, menstruation, in the normal physiological processes such as athleticism, the HSP70 of biosome expresses also and significantly raises, and points out it may play a significant role in these processes.The researcher finds that the autoantibody of Hsps in the generation of a lot of diseases, formation, prognosis process significantly changes, and prompting HSP70 antibody may have vital role in these processes.The HSP70 that it is generally acknowledged induction type is intracellular albumen, but research finds can detect HSP70 and the HSP70 antibody of solubility under normal person's peripheral circulation and some diseases state, intracellular HSP70 can be discharged into the extracellular, causes that body produces HSP70 antibody.Recent research finds that in various rheumatic diseases and autoimmune disease HSP70 can be identified by body, produces autoantibody; Population epidemiology research finds that HSP70 antibody and atherosclerotic progress and the order of severity are closely related.So set up the method that detects fast, reliably HSP70 antibody in the blood, important means will be provided for the monitoring of various diseases.The method that detects now HSP70 antibody mainly contains Western Blotting, Elisa method.The whole bag of tricks corroborates each other, and has again simultaneously the impact of detection sensitivity, professional, installations and facilities condition and cost.At present, the Enzyme-linked Immunosorbent Assay determining adsorption (Elisa) take the serological reaction principle as the basis is the method that is widely accepted and promotes.
Enzyme Linked Immunoadsorbent Assay (enzyme linked immunosorbent assay, ELISA) technology is to carry out at present the body fluid bioprotein to detect method the most commonly used, compare with other detection methods, the ELISA detection method has that cost is low, checkout facility equipment simply, characteristics rapidly and efficiently, and in the middle of updating, Salmonella, indirect ELISA, monoclonal antibody body ELISA have been arranged, microwave ELISA, double-antibodies sandwich ELISA.But the ELISA method is only applicable at present the laboratory and detects, and can not be applicable to Site Detection.
Stress be body in to living environment in the multiple unfavorable factor procedure of adaptation, uneven body and mind tense situation and the reaction thereof that causes between the requirement in reality or the cognition and adaptation and the handling ability.Stress close ties be arranged with people's health, this contact is two-way: stress affect on the one hand people's health, on the other hand a people's health status also can affect psychological stress reaction under the stressed condition intensity and to stress tolerance.The psychological stress of appropriateness is the necessary condition that the people grows up and develops, the suitable physiology that stress help to keep the people, psychology and society function, to cope with challenges both can cause our anxiety, fatigue, worries and painful, can bring the pleasure of success, light and happy for us again.But long-term, surpass that the people adapts to and the health that stress damage the people of adaptibility to response, this be psychological stress to the negative influence of health, be mainly manifested in following three aspects.
At first, the psychology that psychological stress causes and physiological reaction can be indicated in the form of sings and symptoms clinical, become that people are uncomfortable, weakness and the root of mental suffering and the reason that Medical need helps; Secondly, psychological stress can increase the weight of existing phrenoblabia and physical disease, or makes these palindromia; At last, psychological stress can cause the sensitization to disease, and causes new phrenoblabia and physical disease under the acting in conjunction of other factors.
The laboratory early-stage Study finds, long-term high strength stress, body HSP70 antibody horizontal raises, HSP70 antibody can be used as stress biomarker, can detect the physical stress level, the generation of prevention stress damage is used for physical stress load health risk assessment.
Summary of the invention
The present invention seeks to overcome the deficiency in the existing HSP70 antibody test technology, a kind of human blood HSP70 antibody colloidal gold test strip is provided.
The present invention also aims to provide the preparation method of above-mentioned test strip.
A kind of human blood HSP70 antibody colloidal gold mark detection test paper bar, this test strips comprise that being located at the sample pad 2, the gold that link to each other successively on the fixed head 1 marks pad 3, nitrocellulose filter 4 and thieving paper 9; Be provided with 5,6,7 and nature controlling line antibody 8 of three Parallel testing line antigens at nitrocellulose filter 4; Be provided with gold mark antigen at gold mark pad 3.
Fixed head 1 is for having the PVC sheet material of pressure sensitive adhesive, and sample pad 2 is glass fibre membrane, and gold mark pad 3 is the dacron film.
The golden antigen of marking is colour developing antigen, and detection line antigen is capture antigen, and colour developing antigen is combined with capture antigen energy while and HSP 70 antibody and is formed double antigens sandwich; Colour developing antigen and capture antigen are the HSP 70 albumen that RT-PCR is expressed;
Gold mark antigen particle diameter is the colloid gold label of 20nm-60nm.
Nature controlling line antibody is the anti-human HSP70 polyclonal antibody of rabbit.
A kind of preparation method of human blood HSP70 antibody colloidal gold mark detection test paper bar claimed in claim 1 is characterized in that, may further comprise the steps:
(1) prepares the collaurum that uniform grading is 20nm-60nm with conventional trisodium citrate reduction method;
(2) regulating collaurum pH is 8.5-9, then adds colour developing antigen and its reaction, and the concentration of the antigen that wherein develops the color is 0.02mg/ml, centrifugal must the precipitation, and resuspended precipitation gets golden mark antigenic solution, and gold is marked the antigenic solution spray printing at the dacron film, dries;
(3) capture antigen being become concentration with diluted is the capture antigen solution of 1mg/ml, with the ink-jet instrument capture antigen solution is drawn three Parallel testing lines, vacuum drying in the nitrocellulose filter spray;
(4) the anti-human HSP70 Anti-TNF-α of rabbit body and function diluted being become concentration is the anti-human HSP70 Anti-TNF-α of the rabbit liquid solution of 0.5mg/ml, with spray film instrument antibody is sprayed line and forms nature controlling line at nitrocellulose filter, vacuum drying;
(5) sequentially stick on the fixed head slitting, the polybag of packing into, dry packing by sample pad, gold mark pad, nitrocellulose membrane, thieving paper.
Centrifugal method is: reactant liquor precipitates with 1wt% bovine serum albumin solution resuspension, 2 times repeatedly in the centrifugal 30min of 5000xg; Wherein 1wt% bovine serum albumin(BSA) (BSA) solution is the Tris-HCl buffer solution of 10mM, and pH is 8.5, contains the bovine serum albumin(BSA) of 1wt%.
The solution of resuspended precipitation is in the step (2): the Tris-HCl buffer solution of 20mM, and pH is 8.5, contains the trehalose of 30wt%, the bovine serum albumin(BSA) of 1wt%, the casein of 1wt %, volumetric concentration are 0.01% Tween-20(Tween-20), the NaN3 of 0.02 wt %.
State the phosphate buffered solution that dilution is 10mM, pH is 7.6, wherein contains the trehalose of 10wt%, and volumetric concentration is 3% methyl alcohol.
Beneficial effect of the present invention is: be used for human blood HSP70 antibody test, detection sensitivity is high, and accuracy is strong, cost is low; With regard to the energy sentence read result, have good stability and repeated, easy and simple to handle in the easy observe and decide of result, 20min, need not special instruments and equipment, basic hospital and experimental study promotion and application suit to detect at the scene.
Description of drawings
Fig. 1 is the structural representation of test strips; Wherein each label is: the 1-fixed head, and the 2-sample pad, 3-is the collaurum pad, 4-is nitrocellulose filter, 5-the first detection line antigen, 6-the second detection line antigen, 7-the 3rd detection line antigen, 8-is nature controlling line, 9-thieving paper.
Fig. 2 is that the test strips test result is judged synoptic diagram.
Fig. 3 is the SDS-PAGE figure of HSP70 albumen behind the protein induced Expression and purification of HSP70; Each band is respectively: the not purified whole bacterial protein of 1-abduction delivering; 2,3,4 be respectively Ni post affinity purification HSP70 albumen, the target protein of HSP70 shown in the arrow.
Embodiment
The preparation of test strips is carried out in the following manner
1. collaurum preparation: with HAuC1
4Be mixed with the aqueous solution that concentration is 0.01wt%, get 100ml and be heated to and boil.Stir the lower 1wt% trisodium citrate (C that accurately adds 2ml
6H
5Na
3O
72H
2O) aqueous solution.Continue heating and boil 15min.Can be observed flaxen aqueous solution of chloraurate this moment and after sodium citrate adds, become very soon grey, continuous and change into black, be stable into gradually subsequently redness.Return to original volume with distilled water after being cooled to room temperature.
2. the analysis of collaurum quality: adopt multi-functional microplate reader that collaurum is carried out wavelength at 400～800nm scope densitometric scan, obtain its maximum absorption wavelength and half-peak breadth.Judge particle size according to maximum absorption wavelength, judge the uniform particle diameter degree according to half-peak breadth.
3. antigen gold mark develops the color: use K
2CO
3Adjust the pH value to 8.5 of collaurum, in the 100ml colloidal gold solution, drip colour developing antigen 2mg, stir 5min, the static placement of lucifuge 30min adds 25ml 5wt% BSA solution (with the Tris-HCl buffer solution of 10mM, pH is 8.5 preparations) again, more than the sealing 2h, the centrifugal 30min of reactant liquor 5000xg, precipitation suspends with the 1wt% BSA solution weight of 125ml, repeatedly processes 2 times, the precipitation resuspended liquid of 1ml (the Tris-HCl buffer solution of 20mM, pH is 8.5, wherein contains the trehalose of 30wt%, the BSA of 1wt%, the casein of 1wt%, volumetric concentration is 0.01% Tween-20, the NaN3 of 0.02wt%) resuspended precipitation, obtain gold mark antigen.
4. the preparation of gold mark pad: gold is marked antigen with spraying film instrument spray printing on polyester fiber element film, and 2 μ l/cm is optimum, drying.
5. the preparation of detection line: get the HSP70 albumen capture antigen (can form sandwich pairing with colour developing antigen) of purifying as detection line antigen, with the dilution (phosphate buffered solution of 10mM, pH is 7.6, wherein contains the trehalose of 10wt%, and volumetric concentration is 3% methyl alcohol) be diluted to the solution of 1mg/ml concentration, on nitrocellulose filter, antigen is sprayed line with the ink-jet instrument, spray a line every 0.5cm, spray altogether three parallel lines, form detection line, the spray line is optimum with 1 μ l/cm, vacuum drying.
6. the preparation of nature controlling line: with the Anti-TNF-α body and function dilution (phosphate buffered solution of 10mM of the anti-human HSP70 of rabbit, pH is 7.6, the trehalose that wherein contains 10wt%, volumetric concentration is 3% methyl alcohol) to be diluted to concentration be 0.5mg/ml solution, with spray film instrument antibody is sprayed line and form nature controlling line at nitrocellulose filter, 1 μ l/cm is optimum, vacuum drying.
7. the assembling of test strips: on the PVC plate 1 that sample pad 2, gold mark pad 3, nitrocellulose filter 4 and thieving paper 9 are pasted in order at pressure sensitive adhesive, as shown in Figure 1, contain 3 detection lines 5,6,7 and nature controlling line 8 of capture antigen on the nitrocellulose filter 4; To stick good large plate and cut into the wide test strips of 4mm, be packaged in the closed container that contains drying agent room temperature preservation.
8. detect and result's judgement: test strips is kept flat, and sample 100ul drips on sample pad, and observe the testing result interpretation during 20min: detection line shows 1 band, shows feminine gender; Detection line shows 2 bands, shows the weak positive; Detection line shows 3 bands, shows strong positive; Above result judges and must show band as the basis with nature controlling line that if nature controlling line, shows that test strips lost efficacy without the colour developing band, the result judges invalid, redeterminates, as shown in Figure 2.
The test strips of embodiment 1 preparation is to the detection of HSP70 antibody in the human serum
Adopt the HSP70 antibody ELISA detection kit to detect crowd's blood sample, according to testing result, from normal person's blood sample, moderate Stress intensity human blood sample and height are chosen respectively in stress the crowd and are represented each 5 parts of HSP70 negative antibody, the weak positive and strong positive blood samples, 15 parts of blood samples are drawn the plasma sample of 100 μ l with pipettor, are added on the sample pad of colloidal gold strip the beginning timing, the wait red stripes occurs, judged result in the time of 20 minutes.
The result judges:
When nature controlling line occurs, when a line appears in detection line, show the HSP70 negative antibody, show that the physical stress level is normal.
When nature controlling line occurs, when two-lines appears in detection line, show that HSP70 antibody is weak positive, show that body is in time strong stress situation;
When nature controlling line occurs, when three lines appear in detection line, show HSP70 antibody strong positive, show that body was in strong stress situation;
If nature controlling line, shows that test strips lost efficacy without the colour developing band;
The result: 15 test strips nature controlling lines all occur, 15 parts of blood examination results with predict the outcome equally, 100% all meets, and shows that test strips is accurately and reliably.
The preparation purifying of HSP70 proteantigen is as follows:
(1) design pcr amplification primer makes up prokaryotic expression carrier, wherein:
Upstream primer is: 5 ' CGAATTCATGGCCAAGAAAACAGCG3 '; Downstream primer is: 5 ' CCCAAGCTTCTAATCCACCTCCTCGAT3 ', pcr amplification HSP70 gene.
The pcr amplification condition is:
Phase one: temperature: 95 ℃, the time: 3min;
Subordinate phase: temperature: 95 ℃, time: 30s, temperature: 56 ℃, time: 60s, temperature: 72 ℃, time 60s, more than carry out 30 circulations;
Phase III: temperature: 72 ℃, time: 10min.
The PCR product purification reclaims for subsequent use by EcoRI and Hind III double digestion by glue recovery kit;
(2) make up the prokaryotic expression bacterial strain
Expression vector PET32a reclaims the kit recovery through EcoRI and Hind III double digestion by glue, reclaiming fragment with the HSP70PCR product is connected, be converted in the BL21 competence bacterial strain, be coated with flat board, in incubator, cultivate 16h, choose the clone, in shaking tube, expand, carry plasmid enzyme restriction and identify that positive bacteria is frozen, as the HSP70 expression strain.
(3) abduction delivering and purifying HSP70 albumen
The HSP70 expression strain is 37 ℃ of joltings in the LB nutrient culture media, when being expanded to OD value 0.5, add IPTG 1.0U/ml, induce 4h, centrifugal collection thalline, ultrasonication in the PB damping fluid, centrifugal collection supernatant.The HP HISTrip of the supernatant GE company purification column of collecting is carried out purifying.
Purification condition is:
Affine damping fluid (Buffer): the phosphate buffer of 20mM, sodium chloride concentration are 500mM, and pH is 7.4.
The phosphate buffer of wash-out Buffer:20mM, sodium chloride concentration are 500mM, and imidazole concentration is 500mM, and pH is 7.4.
Sample thief 10ml is through 0.45 μ m membrane filtration; Pillar (Histrap 5ml) is with 10 column volumes of affine Buffer balance, and flow velocity is 5ml/min, loading 10ml, wash 5 column volumes through affine Buffer again, use wash-out Buffer, 100ml linear gradient elution target protein, collect target peak, carry out Purity.
4) identify purity of protein with the SDS-PAGE electrophoresis
The albumen of wash-out purifying is mixed with sample-loading buffer, 100 ℃ of heating 5min sex change, the centrifugal 10min of 1000rpm gets supernatant 10ul loading, and the SDS-PAGE gum concentration is 12.5wt%, deposition condition, 60V, 30min, 120V, 80min; After electrophoresis was complete, glue carried out coomassie brilliant blue R250 dyeing, and after the decolouring, glue scans and graphical analysis, the result: restructuring HSP70 purity of protein＞and more than 95%, as shown in Figure 3.
5) eluent is packed in the bag filter, dialysed overnight in the PBS liquid is changed liquid therebetween three times.Liquid subpackage in the bag filter is entered in the freeze drying pipe, namely make the HIS-HSP70 recombinant protein, total protein concentration is 1mg in every pipe, and freeze drying is powdered, put into-70 ℃ for subsequent use.
The anti-human HSP70 polyclonal antibody preparation of rabbit
1) select the Healthy female rabbit, the HIS-HSP70 recombinant protein made from embodiment 2 is as the immunogen immune animal; Immune programme for children is: be that the 2mg/ml proteantigen mixes fully with the Freund's complete adjuvant of 0.5ml with the concentration of 0.5ml, and at the subcutaneous multi-point injection in rabbit back, every some 0.2ml, lower limb groin 0.2ml;
2) after one month, be that the Freund's complete adjuvant of 2mg/ml proteantigen and 0.5ml mixes fully with the concentration of 0.5ml, at the subcutaneous multi-point injection in rabbit back, every some 0.2ml, lower limb groin 0.2ml;
3) after another month, be that 2mg/ml proteantigen ear vein is injected with the concentration of 0.5ml;
4) again after 7 days, the arteria carotis bloodletting gathers, separation of serum, and adding concentration is that 1wt% thimerosal aqueous solution is anticorrosion ,-70 degree are preserved,
5) adopt ammonium sulfate precipitation method purifying HSP70 polyclonal antibody, obtain the anti-human HSP70 polyclonal antibody of rabbit.
Claims (9)
1. a human blood HSP70 antibody colloidal gold mark detection test paper bar is characterized in that: comprise being located at sample pad (2), gold mark pad (3), nitrocellulose filter (4) and the thieving paper (9) that links to each other successively on the fixed head (1); Be provided with three the first parallel detection line antigens (5), the second detection line antigen (6), the 3rd detection line antigen (7) and nature controlling line antibody (8) at nitrocellulose filter (4); Be provided with gold mark antigen at gold mark pad (3).
2. human blood HSP70 antibody colloidal gold mark detection test paper bar according to claim 1 is characterized in that: described fixed head (1) is for having the PVC sheet material of pressure sensitive adhesive, and sample pad (2) be glass fibre membrane, and it is the dacron film that the gold mark fills up (3).
3. human blood HSP70 antibody colloidal gold mark detection test paper bar according to claim 1, it is characterized in that: described gold mark antigen is colour developing antigen, detection line antigen is capture antigen, and colour developing antigen is combined with capture antigen energy while and HSP 70 antibody and is formed double antigens sandwich; Colour developing antigen and capture antigen are the HSP 70 albumen that RT-PCR is expressed.
4. human blood HSP70 antibody colloidal gold mark detection test paper bar according to claim 1 is characterized in that: described gold mark antigen particle diameter is the colloid gold label of 20nm-60nm.
5. human blood HSP70 antibody colloidal gold mark detection test paper bar according to claim 1, it is characterized in that: described nature controlling line antibody is the anti-human HSP70 polyclonal antibody of rabbit.
6. the preparation method of a human blood HSP70 antibody colloidal gold mark detection test paper bar claimed in claim 1 is characterized in that, may further comprise the steps:
(1) prepares the collaurum that uniform grading is 20nm-60nm with conventional trisodium citrate reduction method;
(2) regulating collaurum pH is 8.5-9, then adds colour developing antigen and its reaction, and the concentration of the antigen that wherein develops the color is 0.02mg/ml, centrifugal must the precipitation, and resuspended precipitation gets golden mark antigenic solution, and gold is marked the antigenic solution spray printing at the dacron film, dries;
(3) capture antigen being become concentration with diluted is the capture antigen solution of 1mg/ml, with the ink-jet instrument capture antigen solution is drawn three Parallel testing lines, vacuum drying in the nitrocellulose filter spray;
(4) the anti-human HSP70 Anti-TNF-α of rabbit body and function diluted being become concentration is the anti-human HSP70 Anti-TNF-α of the rabbit liquid solution of 0.5mg/ml, with spray film instrument antibody is sprayed line and forms nature controlling line at nitrocellulose filter, vacuum drying;
(5) sequentially stick on the fixed head slitting, the polybag of packing into, dry packing by sample pad, gold mark pad, nitrocellulose membrane, thieving paper.
7. preparation method according to claim 6 is characterized in that, centrifugal method is: reactant liquor precipitates with 1wt% bovine serum albumin solution resuspension, 2 times repeatedly in the centrifugal 30min of 5000xg; Wherein the 1wt% bovine serum albumin solution is the Tris-HCl buffer solution of 10mM, and pH is 8.5, contains the bovine serum albumin(BSA) of 1wt%.
8. preparation method according to claim 6, it is characterized in that, the solution of resuspended precipitation is in the step (2): the Tris-HCl buffer solution of 20mM, pH is 8.5, the trehalose that contains 30wt%, the bovine serum albumin(BSA) of 1wt%, the casein of 1wt %, volumetric concentration is 0.01% Tween-20, the NaN3 of 0.02 wt %.
9. preparation method according to claim 6 is characterized in that, described dilution is the phosphate buffered solution of 10mM, and pH is 7.6, wherein contains the trehalose of 10wt%, and volumetric concentration is 3% methyl alcohol.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210365239XA CN102879584A (en) | 2012-09-26 | 2012-09-26 | Human blood HSP70 antibody colloidal gold-labeled detection test strip and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210365239XA CN102879584A (en) | 2012-09-26 | 2012-09-26 | Human blood HSP70 antibody colloidal gold-labeled detection test strip and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102879584A true CN102879584A (en) | 2013-01-16 |
Family
ID=47480983
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210365239XA Pending CN102879584A (en) | 2012-09-26 | 2012-09-26 | Human blood HSP70 antibody colloidal gold-labeled detection test strip and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102879584A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113588960A (en) * | 2020-11-04 | 2021-11-02 | 北京北方生物技术研究所有限公司 | Immunochromatography detection test strip by ratio fluorescence method and detection method thereof |
| CN115575623A (en) * | 2022-12-06 | 2023-01-06 | 深圳市卓润生物科技有限公司 | Colloidal gold and preparation method and application thereof |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1547024A (en) * | 2003-12-03 | 2004-11-17 | 华南农业大学 | Fast and semiquantitative detection method for Clenbuterol |
| CN1886425A (en) * | 2003-09-25 | 2006-12-27 | 弗塞沃洛德·I·基塞莱夫 | Methods, kits, and compositions for the development and use of monoclonal antibodies specific to antigens traditionally of low immunogenicity |
| CN101105498A (en) * | 2007-08-08 | 2008-01-16 | 中国人民解放军军事医学科学院卫生学环境医学研究所 | Human, rat, mouse HSP70 double antibody sandwich method detection reagent kit |
| WO2009023846A2 (en) * | 2007-08-15 | 2009-02-19 | The Research Foundation Of State University Of New York | Methods for heat shock protein dependent cancer treatment |
| CN101498730A (en) * | 2009-03-16 | 2009-08-05 | 深圳市菲鹏生物股份有限公司 | Improved double-antigen sandwiching immunity detection method |
| CN101833003A (en) * | 2010-05-26 | 2010-09-15 | 吉林大学 | Vesicular stomatitis virus antibody colloidal goldtest paper tape and preparation method thereof |
| CN101881770A (en) * | 2009-05-08 | 2010-11-10 | 青岛农业大学 | Method for preparing porcine circovirus type 2 colloidal gold antibody fast test strip |
| WO2011062469A2 (en) * | 2009-11-23 | 2011-05-26 | Universidad Nacional Autónoma de México | Diagnostic method for detecting acute kidney injury using heat shock protein 72 as a sensitive biomarker |
| EP2330424A1 (en) * | 2008-07-02 | 2011-06-08 | Aposcience AG | COPD diagnosis |
| CN102507950A (en) * | 2011-11-23 | 2012-06-20 | 中国人民解放军军事医学科学院基础医学研究所 | Plasma HSP70 (Heat Shock Protein 70) antibody detection kit |
| CN102565407A (en) * | 2011-12-22 | 2012-07-11 | 正元盛邦(天津)生物科技有限公司 | Method for semi-quantitative diagnosis of clenbuterol hydrochloride through double-indicating line immunochromatography |
| CN102608313A (en) * | 2012-02-27 | 2012-07-25 | 中国疾病预防控制中心病毒病预防控制所 | Anti-hepatitis A virus IgM (immunoglobulin M) AlphaLISA detection kit |
-
2012
- 2012-09-26 CN CN201210365239XA patent/CN102879584A/en active Pending
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1886425A (en) * | 2003-09-25 | 2006-12-27 | 弗塞沃洛德·I·基塞莱夫 | Methods, kits, and compositions for the development and use of monoclonal antibodies specific to antigens traditionally of low immunogenicity |
| CN1547024A (en) * | 2003-12-03 | 2004-11-17 | 华南农业大学 | Fast and semiquantitative detection method for Clenbuterol |
| CN101105498A (en) * | 2007-08-08 | 2008-01-16 | 中国人民解放军军事医学科学院卫生学环境医学研究所 | Human, rat, mouse HSP70 double antibody sandwich method detection reagent kit |
| WO2009023846A2 (en) * | 2007-08-15 | 2009-02-19 | The Research Foundation Of State University Of New York | Methods for heat shock protein dependent cancer treatment |
| EP2330424A1 (en) * | 2008-07-02 | 2011-06-08 | Aposcience AG | COPD diagnosis |
| CN101498730A (en) * | 2009-03-16 | 2009-08-05 | 深圳市菲鹏生物股份有限公司 | Improved double-antigen sandwiching immunity detection method |
| CN101881770A (en) * | 2009-05-08 | 2010-11-10 | 青岛农业大学 | Method for preparing porcine circovirus type 2 colloidal gold antibody fast test strip |
| WO2011062469A2 (en) * | 2009-11-23 | 2011-05-26 | Universidad Nacional Autónoma de México | Diagnostic method for detecting acute kidney injury using heat shock protein 72 as a sensitive biomarker |
| CN101833003A (en) * | 2010-05-26 | 2010-09-15 | 吉林大学 | Vesicular stomatitis virus antibody colloidal goldtest paper tape and preparation method thereof |
| CN102507950A (en) * | 2011-11-23 | 2012-06-20 | 中国人民解放军军事医学科学院基础医学研究所 | Plasma HSP70 (Heat Shock Protein 70) antibody detection kit |
| CN102565407A (en) * | 2011-12-22 | 2012-07-11 | 正元盛邦(天津)生物科技有限公司 | Method for semi-quantitative diagnosis of clenbuterol hydrochloride through double-indicating line immunochromatography |
| CN102608313A (en) * | 2012-02-27 | 2012-07-25 | 中国疾病预防控制中心病毒病预防控制所 | Anti-hepatitis A virus IgM (immunoglobulin M) AlphaLISA detection kit |
Non-Patent Citations (3)
| Title |
|---|
| 刘志宗等: "猪肺炎支原体热休克蛋白HSP70的研究进展", 《畜牧兽医科技信息》, no. 3, 31 December 2009 (2009-12-31), pages 10 - 11 * |
| 张北奇等: "不同应激刺激后大鼠心脏、肝脏热休克蛋白(HSP70)的产生规律", 《河北医药》, vol. 25, no. 8, 31 August 2003 (2003-08-31), pages 563 - 565 * |
| 王彦中等: "热休克蛋白HSP70和gp96增强乙肝DNA疫苗的细胞和体液免疫应答", 《生物工程学报》, vol. 27, no. 5, 25 May 2011 (2011-05-25), pages 790 - 798 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113588960A (en) * | 2020-11-04 | 2021-11-02 | 北京北方生物技术研究所有限公司 | Immunochromatography detection test strip by ratio fluorescence method and detection method thereof |
| CN115575623A (en) * | 2022-12-06 | 2023-01-06 | 深圳市卓润生物科技有限公司 | Colloidal gold and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104198703B (en) | People's mycoplasma pneumoniae gold label silver stain immunochromatographytest test kit and its preparation method and application | |
| CN105527437B (en) | A kind of detection kit and its application | |
| CN105695420B (en) | Mouse bone marrow cells hybridoma cell strain and its monoclonal antibody and application generated | |
| CN109781980A (en) | African swine fever virus rapid detection card and its application | |
| CN105675873B (en) | A kind of detection kit and its application | |
| CN102539782A (en) | Immune chromatography test strip for detecting cystic echinococcosis and alveolar echinococcosis and preparation method | |
| CN101825634A (en) | Reagent strip for joint detection of syphilis specific IgM and IgG antibodies and preparation method thereof | |
| CN101858914B (en) | Reagent strip for testing syphilis specific total antibodies through gold immunochromatographic assay and preparation method thereof | |
| CN110058019A (en) | A kind of hog cholera antibody blocking Test paper | |
| CN109187968A (en) | A kind of swine fever virus and the bigeminy gold mark detection test paper of porcine pseudorabies virus and preparation method thereof | |
| CN105753981B (en) | The immune chromatography reagent kit of anti-human Respiratory Syncytial Virus(RSV) N protein antibody and the application antibody | |
| CN103823057B (en) | Total antibody colloidal gold fast diagnose test paper bar of one boar HEV and preparation method thereof | |
| CN102879584A (en) | Human blood HSP70 antibody colloidal gold-labeled detection test strip and preparation method thereof | |
| CN101858916A (en) | Reagent strip for testing syphilis specific IgM antibodies through gold immunochromatographic assay and preparation method thereof | |
| CN103278640A (en) | Anti-BP180 NC16A IgE antibody ELISA kit and detection method | |
| CN101858915A (en) | Reagent strip for testing syphilis specific IgG antibodies through gold immunochromatographic assay and preparation method thereof | |
| CN106990247A (en) | Colloidal gold strip of 1 type and 3 type duck hepatitis A virus and preparation method thereof is detected simultaneously | |
| CN102109523A (en) | Hepatitis C virus IgG and IgM antibody joint detection kit and preparation method thereof | |
| CN109406788A (en) | A kind of monoclonal antibody and its application | |
| CN105891469B (en) | A kind of Portunus trituberculatus Miers reovirus test strip and preparation method thereof | |
| CN102890156A (en) | Gold-labeled detection test strip of body fluid HSP70 gels and preparation method of gold-labeled detection test strip | |
| CN107557344A (en) | One plant of expression CSFV E 2 protein monoclonal antibody hybridoma cell line CSFV 3H3G6 and antibody and kit | |
| CN102608330A (en) | Immunochromatographic strip for detecting alveolar echinococcosis and preparation method thereof | |
| CN110144330A (en) | Hybridoma, monoclonal antibody and the application of anti-PEDV monoclonal antibody can be secreted | |
| CN107478838A (en) | A kind of colloidal gold immuno-chromatography test paper strip for detecting people and the anti-RVF antiviral antibodies of animal blood serum |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20130116 |