CN204287199U - H-FABP immunochromatographiassay assay quantitative detection test paper - Google Patents
H-FABP immunochromatographiassay assay quantitative detection test paper Download PDFInfo
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- CN204287199U CN204287199U CN201420645367.4U CN201420645367U CN204287199U CN 204287199 U CN204287199 U CN 204287199U CN 201420645367 U CN201420645367 U CN 201420645367U CN 204287199 U CN204287199 U CN 204287199U
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Abstract
The utility model discloses a kind of H-FABP immunochromatographiassay assay quantitative detection test paper, comprise base plate, sample pad, fluorescent marker pad, cellulose nitrate and adsorptive pads; Described test strips is overlapped successively to be pasted onto on base plate formed by sample pad, fluorescent marker pad, cellulose nitrate, adsorptive pads.Described fluorescent marker pad contains marked by streptavidin fluorescin and biotin labeled heart fatty acid binding protein monoclonal antibody; Described nitrocellulose membrane has detection line and nature controlling line, detection line is by heart fatty acid binding protein monoclonal antibody, and it has different identification epi-positions from above-mentioned biotin labeled heart fatty acid binding protein monoclonal antibody.The utility model is (as: chemoluminescence method/Immune-enhancing effect turbidimetry/colloidal gold method) compared with the method for detection H-FABP common at present, not only substantially reduces detection time, also improves detection sensitivity simultaneously.
Description
Technical field
The utility model belongs to field of immunodetection, is specifically related to a kind of high sensitivity H-FABP immunochromatographiassay assay quantitative detection test paper.
Background technology
Fatty acid binding protein (Fatty acid binding protein, FABP) is the small molecular protein family that several functions is identical, is mainly present in the cell cytoplasm of a large amount of power consumption, plays a part to transport free fatty acid in cell in fat metabolism.Be also called H-FABP (heart-type fatty acid-binding protein prior to the FABP found in cardiac muscle, be called for short H-FABP), that a kind of low-molecular-weight cardiac muscle cell starches non-enzyme albumen, molecular weight is 14500D, be made up of 132 amino acid residues, its molecular structure and immunological characteristic are different from the FABP of other types, have Cardiac-specific.
The mark of current clinical conventional diagnosis AMI has CK-MB, Mb, cardiac troponin (cardiac troponin, cTn) etc., and existing Applications of Cardiac Markers often can not be taken into account in sensitivity and specificity.CTnI, cTnT are the most special Applications of Cardiac Markers found so far, but cTn raises relatively evening the long period of continuing, and its value in AMI early diagnosis and reflecting myocardium again infarct is still limited.The shortcoming of above Applications of Cardiac Markers makes the early diagnosis of AMI become more difficult.Along with the discovery of H-FABP and the introducing at AMI diagnostic field thereof, this situation may improve.As the marker protein of a kind of new AMI, H-FABP day by day comes into one's own and pays close attention in recent years.
H-FABP can find super anxious phase AMI patient early.Need the patient of emergency hospitalization, coronarography, interventional therapy to have very high susceptibility for identifying, early diagnosis and the Index for diagnosis of the patient especially in those pectoralgias morbidity 6h have very great help.H-FABP detects together with combining with traditional label cTnI, CK-MB, early detection H-FABP, convalescence detects cTnI, be conducive to having filtered out the high risk patient of cardiac disorder, not only conveniently take positive therapeutic strategy, also can save patient's spending and lab resources, perhaps H-FABP and cTnI rationally complementary monitoring is the ideal chose of a kind of clinical diagnosis and treatment ischemic heart disease in the future.
The common method of current detection H-FABP has:
1 Latex Agglutination
Latex Agglutination is clinical comparatively conventional serological method, belongs to indirectly agglutination test.The anti-human H-FABP antibody sensitized of emulsion reagent purifying, can and patients serum in H-FABP there is specific reaction, present agglutinating particle clearly in several minutes, occur that aggegation person is for positive, do not occur that aggegation person is for feminine gender.The method is simple to operate, quick, and susceptibility, specificity are higher.But be subject to the interference of the factor such as complement, rheumatoid factor (RF), produce false positive results.Therefore, in order to improve the accuracy of result, tackling sample to be measured during detection and carrying out pre-service, to remove disturbing factor.
2 Immunity transmission turbidity
Immunity transmission turbidity is that detection method is commonly used in laboratory.Immunity transmission turbidity is also a kind of immunoprecipitation assay of trace.Itself and rate nephelometry are unlike the content reflecting determined antigen with the light quantity measured through solution.When light therethrough reaction system, the antigenantibody complex in solution can be absorbed light and be reflected, and transmitted light is reduced.Immune complex is more, and the light of absorption is more, and transmitted light is fewer, and the available absorbance of this change represents.If antibody amount is fixed, survey absorbance and be directly proportional to the amount of immune complex, be also directly proportional to the amount of determined antigen.Compare with the antigen standard of a series of concentration known, namely can measure examined object content.Nanjing Norman Biological Technology Co., Ltd. H-FABP measures H-FABP content set up by reagent calibration curve by reference to calibration object, can measure the content of H-FABP in serum on automatic clinical chemistry analyzer fast.
3 immunolabelling techniques
The immune labeled method that immunolabelling technique is used for H-FABP mensuration has radioimmunology, enzyme immunoassay, gold-marking immunity method, fluorescent mark immunity method etc.The sensitivity of IRNA method and specificity high, and do not need separation and purification blood plasma, fast more practical than RIA method, but need 5 ~ 36 hours consuming time, be not suitable for automatic analysis system.Domestic also have part Related product, the H-FABP of auspicious Lay bioengineering (Shenzhen) company limited detects reagent and utilizes two-way effluent immunization, can realize detecting H-FABP concentration in the scope quantification of 2ng/mL-60ng/mL by the change measuring golden labeling antibody optical density on detection paper line and nature controlling line.
Biotin-avidin system (biotinavidin system, BAS) is a kind of new bio reaction amplification system.Along with the appearance of various biotin derivative, BAS is widely used in each field of medical science very soon.This system is applied to SABC, enzyme linked immunological, fluorescence immunoassay, in the detection techniques such as radio-immunity, the susceptibility of above technical method, specificity and stability can be improved significantly, make method easier, contribute to clinical quick diagnosis, and be beneficial to and carry out extensive epidemiology survey, become the immunoreactive powerful of research.Because BAS detection system economy is quick, pollute without radiating matter again, do not need complex instrument, fully show the great potential of this system and the possibility of application.
Biotin-avidin system Cleaning Principle: BAS is on the basis of conventional ELISA principle, in conjunction with the height amplification between biotin and Avidin, and a kind of detection system set up.Avidin is a kind of alkaline glycoprotein extracted in ovalbumin, and molecular weight is 68kDa, is made up of 4 subunits, have very high affinity (binding constant is up to 1015M-1) to biology.Biotin is very easily combined with covalent bond with protein (as antibody etc.).Like this, the Avidin molecule combining enzyme and the biotin molecule being combined with specific antibody produce and react, both served multistage amplification, the colour generation due to the catalytic action of enzyme when running into corresponding substrate, reaches the object detecting unknown antigen (or antibody) molecule again.Streptavidin is and the affine a kind of protein have similar biological properties, in Streptavidin molecule the same as Avidin, every bar peptide chain can in conjunction with a biotin, because nearly all material for marking all can combine with Avidin or streptavidin.Utilize biotin-avidin system to develop H-FABP detection examination fast diagnosis reagent and there is no report.
Utility model content
The purpose of this utility model, be to provide a kind of H-FABP immunochromatographiassay assay quantitative detection test paper, it provides high sensitivity H-FABP immunochromatographiassay assay quantitative detection test paper based on double antibody sandwich method, fluorescence immunoassay lateral chromatography and biotin-Streptavidin amplification system.When using the utility model, improve detection sensitivity and detect stability, reducing non-specific binding, be not more than 10 minutes detection time, substantially increase diagnosis efficiency.
The solution that the utility model solves its technical matters is: H-FABP immunochromatographiassay assay quantitative detection test paper, it comprises base plate, described base plate is provided with sample pad successively, fluorescent marker pad, nitrocellulose filter and adsorptive pads, described adsorptive pads and fluorescent marker pad are overlapping to be respectively pressed in behind the two ends of nitrocellulose filter in the formation detection zone, surface of nitrocellulose filter, described sample pad is overlapping to be pressed on fluorescent marker pad, described fluorescent marker pad is fixed with the fluorescin of biotin labeled heart fatty acid binding protein monoclonal antibody and marked by streptavidin, nitrocellulose filter in described detection zone is fixed with the nature controlling line of detection line and the sheep anti-mouse igg polyclonal antibody formation identifying that the monoclonal antibody of H-FABP another one epi-position is formed.
As the further improvement of technique scheme, described fluorescent marker pad comprises the first stacked fluorescent marker pad and the second fluorescent marker pad, one end pad of described first fluorescent marker pad in the below of sample pad, described second overlapping one end being pressed in nitrocellulose filter of fluorescent marker pad.
As the further improvement of technique scheme, in described first fluorescent marker pad, the one end of inserting below sample pad is provided with positioning strip, and the lower surface of described sample pad is provided with the locating slot that can hold positioning strip.
As the further improvement of technique scheme, also comprise and getting stuck, in described base plate, sample pad, fluorescent marker pad, nitrocellulose filter and adsorptive pads are all placed in and get stuck, the position described shell face of getting stuck corresponding to nitrocellulose membrane is provided with view window, and position shell face of getting stuck corresponding to sample pad is provided with well.
As the further improvement of technique scheme, described view window is slot.
As the further improvement of technique scheme, described fluorescin can be the one in green fluorescent protein, phycobniliprotein.
As the further improvement of technique scheme, described sample pad is glass fibre component dry after surfactant damping fluid immersion treatment.
As the further improvement of technique scheme, described base plate is polystyrene component or tygon component.
As the further improvement of technique scheme, described in get stuck and comprise plastics upper casing and plastics lower casing, described plastics upper casing fastens to be formed after on plastics lower casing and gets stuck.
Compared with prior art, tool has the following advantages the utility model:
(1) the utility model is using fluorescin as label, and this label has good stability, and is conducive to improving detecting stability.
(2) the utility model is by utilizing " Streptavidin-biotin amplification system ", improves detection sensitivity, reduces non-specific binding, is conducive to improving kit performance.
(3) detection time utilizing the utility model to carry out Procalcitonin is not more than 10 minutes, and the detection range of linearity is 0.10ng/ml ~ 100.00ng/ml, drastically increases detection efficiency.
(4) the utility model carries out interpretation by fluorescence immunity analyzer to result, can realize robotization, reduces the impact of subjective factor, provides convenient, quick, reliable diagnostic result.
(5) novel the getting stuck of this use is provided with the well that sample enters and the view window supplying observed result, according to analytical instrument result of determination, accurately and reliably.
(6) the utility model is easy to make, and volume is little, be easy to carry.
(7) the utility model testing cost is lower.
(8) the utility model can be mass, and is applicable to clinical quick diagnosis and on-the-spot quick diagnosis; Be easy to preserve, be conducive to grass-roots unit and promote.
Accompanying drawing explanation
In order to be illustrated more clearly in the technical scheme in the utility model embodiment, below the accompanying drawing used required in describing embodiment is briefly described.Obviously, described accompanying drawing is a part of embodiment of the present utility model, instead of whole embodiment, and those skilled in the art, under the prerequisite not paying creative work, can also obtain other design proposals and accompanying drawing according to these accompanying drawings.
Fig. 1 is the structural representation of the utility model embodiment one;
Fig. 2 is the structural representation of the utility model embodiment two;
Fig. 3 is the structural representation of the utility model embodiment three;
Fig. 4 is the structural representation of the utility model embodiment four.
Embodiment
Be clearly and completely described below with reference to embodiment and the accompanying drawing technique effect to design of the present utility model, concrete structure and generation, to understand the purpose of this utility model, characteristic sum effect fully.Obviously; described embodiment is a part of embodiment of the present utility model, instead of whole embodiment, based on embodiment of the present utility model; other embodiments that those skilled in the art obtains under the prerequisite not paying creative work, all belong to the scope of the utility model protection.In addition, all connection/annexations mentioned in literary composition, not singly refer to that component directly connects, and refer to and according to concrete performance, can connect auxiliary by adding or reducing, and form more excellent draw bail.
With reference to Fig. 1, H-FABP immunochromatographiassay assay quantitative detection test paper, it comprises base plate 5, described base plate 5 is provided with sample pad 1 successively, fluorescent marker pad 2, nitrocellulose filter 3 and adsorptive pads 4, described adsorptive pads 4 and fluorescent marker pad 2 are overlapping to be respectively pressed in behind the two ends of nitrocellulose filter 3 in the formation detection zone, surface of nitrocellulose filter 3, fluorescent marker pad 2 is glass fibre membrane, described sample pad 1 is overlapping to be pressed on fluorescent marker pad 2, the fluorescin described fluorescent marker pad 2 being fixed with biotin labeled heart fatty acid binding protein monoclonal antibody (concentration 0.3 ~ 1.5mg/mL) and marked by streptavidin (or Avidin) (uses excitation wavelength 530nm, wavelength of transmitted light 570nm, concentration 0.1 ~ 1.0mg/mL), nitrocellulose filter 3 in described detection zone is fixed with the nature controlling line C (concentration 0.2 ~ 2.0mg/mL) of detection line T (concentration 0.5 ~ 3mg/mL) and the sheep anti-mouse igg polyclonal antibody formation identifying that the monoclonal antibody of H-FABP another one epi-position is formed, nature controlling line C is used for the validity of test strip.
The fluorescin that biotin labeled heart fatty acid binding protein monoclonal antibody and Streptavidin (or Avidin) mark is fixed on fluorescent marker pad 2, the monoclonal antibody and sheep anti-mouse igg polyclonal antibody that have identification H-FABP another one epi-position are fixed on nitrocellulose filter as detection line T and nature controlling line C.When testing sample is added to after in sample pad 1, moved forward by chromatography effect, on sample H-FABP in rat and fluorescent marker pad 2, the H-FABP antibody (Mab-H-FABP*Fluoro) of combined with fluorescent label (fluorescin) reacts and forms compound H-FABP-Mab-H-FABP*Fluoro, compound is reacted when continuing to be advanced past FABP antibody (detection line) nitrocellulose membrane wrapping quilt under chromatography effect, FABP antibody capture formation compound (Mab-H-FABP-β 2-MG-Mab-H-FABP*Fluoro) (detection line) that reaction compound is coated, the reaction signal of detection line is read by fluorescence immunity analyzer, under the effect of excitation source, fluorescent material launches the fluorescence signal of specific wavelength, fluorescence immunity analyzer captures fluorescence signal, the typical curve transformed by signal and set is converted into quantitative value automatically, calculate the concentration of the dirty type fatty acid binding protein of center of a sample, obtain H-FABP testing result.
As the further improvement of technique scheme, with reference to Fig. 3, described fluorescent marker pad 2 comprises the first stacked fluorescent marker pad 20 and the second fluorescent marker pad 21, one end pad of described first fluorescent marker pad 20 in the below of sample pad 1, described second overlapping one end being pressed in nitrocellulose filter 3 of fluorescent marker pad 21.The layering of fluorescent marker pad 2 is arranged, and is convenient to fluorescent marker pad 2 to be arranged between sample pad 1 and nitrocellulose filter 3.
As the further improvement of technique scheme, be provided with positioning strip 22 with reference to one end of inserting below sample pad in the first fluorescent marker pad 20 described in Fig. 3, the lower surface of described sample pad 1 is provided with the locating slot that can hold positioning strip 22.The installation be convenient between fluorescent marker pad 2 and sample pad 1 by positioning strip 22 is fixed.
As the further improvement of technique scheme, with reference to Fig. 2 and Fig. 4, also comprise and getting stuck, described base plate 5, sample pad 1, fluorescent marker pad 2, nitrocellulose filter 3 and adsorptive pads 4 are all placed in and get stuck in 6, the position described 6 shell faces of getting stuck corresponding to nitrocellulose membrane 3 is provided with view window 8, and the position 6 shell faces of getting stuck corresponding to sample pad 1 is provided with well 7.
As the further improvement of technique scheme, described view window 8 is slot.
As the further improvement of technique scheme, described fluorescin can be the one in green fluorescent protein, phycobniliprotein.
As the further improvement of technique scheme, described sample pad 1 is glass fibre component dry after surfactant damping fluid immersion treatment.Sample pad is made up of glass fibre, through surfactant damping fluid immersion treatment, uses after dry.The cutting width of described sample pad 1 is 0.3 ~ 0.5cm.
As the further improvement of technique scheme, described base plate 5 is polystyrene component or tygon component.
As the further improvement of technique scheme, described in get stuck and 6 comprise plastics upper casing 60 and plastics lower casing 61, described plastics upper casing 61 fastens to be formed after on plastics lower casing 61 and gets stuck 6.
More than that better embodiment of the present utility model is illustrated, but the invention is not limited to described embodiment, those of ordinary skill in the art also can make all equivalent modifications or replacement under the prerequisite without prejudice to the utility model spirit, and these equivalent modification or replacement are all included in the application's claim limited range.
Claims (9)
1. H-FABP immunochromatographiassay assay quantitative detection test paper, it is characterized in that: it comprises base plate, described base plate is provided with sample pad successively, fluorescent marker pad, nitrocellulose filter and adsorptive pads, described adsorptive pads and fluorescent marker pad are overlapping to be respectively pressed in behind the two ends of nitrocellulose filter in the formation detection zone, surface of nitrocellulose filter, described sample pad is overlapping to be pressed on fluorescent marker pad, described fluorescent marker pad is fixed with the fluorescin of biotin labeled heart fatty acid binding protein monoclonal antibody and marked by streptavidin, nitrocellulose filter in described detection zone is fixed with the nature controlling line of detection line and the sheep anti-mouse igg polyclonal antibody formation identifying that the monoclonal antibody of H-FABP another one epi-position is formed.
2. H-FABP immunochromatographiassay assay quantitative detection test paper according to claim 1, it is characterized in that: described fluorescent marker pad comprises the first stacked fluorescent marker pad and the second fluorescent marker pad, one end pad of described first fluorescent marker pad in the below of sample pad, described second overlapping one end being pressed in nitrocellulose filter of fluorescent marker pad.
3. H-FABP immunochromatographiassay assay quantitative detection test paper according to claim 2, it is characterized in that: in described first fluorescent marker pad, the one end of inserting below sample pad is provided with positioning strip, and the lower surface of described sample pad is provided with the locating slot that can hold positioning strip.
4. the H-FABP immunochromatographiassay assay quantitative detection test paper according to any one of claims 1 to 3, it is characterized in that: also comprise and getting stuck, in described base plate, sample pad, fluorescent marker pad, nitrocellulose filter and adsorptive pads are all placed in and get stuck, the position described shell face of getting stuck corresponding to nitrocellulose membrane is provided with view window, and position shell face of getting stuck corresponding to sample pad is provided with well.
5. H-FABP immunochromatographiassay assay quantitative detection test paper according to claim 4, is characterized in that: described view window is slot.
6. H-FABP immunochromatographiassay assay quantitative detection test paper according to claim 4, is characterized in that: described fluorescin can be the one in green fluorescent protein, phycobniliprotein.
7. H-FABP immunochromatographiassay assay quantitative detection test paper according to claim 4, is characterized in that: described sample pad is glass fibre component dry after surfactant damping fluid immersion treatment.
8. H-FABP immunochromatographiassay assay quantitative detection test paper according to claim 4, is characterized in that: described base plate is polystyrene component or tygon component.
9. H-FABP immunochromatographiassay assay quantitative detection test paper according to claim 4, is characterized in that: described in get stuck and comprise plastics upper casing and plastics lower casing, described plastics upper casing fastens to be formed after on plastics lower casing and gets stuck.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105527445A (en) * | 2015-12-30 | 2016-04-27 | 天津诺星生物医药科技有限公司 | Acute myocardial infarction detection system |
| CN107505459A (en) * | 2017-07-03 | 2017-12-22 | 广州瑞博奥生物科技有限公司 | Quantitatively detect people H FABP time-resolved fluoroimmunoassay chromatograph test strip, kit and preparation method thereof |
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2014
- 2014-10-28 CN CN201420645367.4U patent/CN204287199U/en active Active
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105527445A (en) * | 2015-12-30 | 2016-04-27 | 天津诺星生物医药科技有限公司 | Acute myocardial infarction detection system |
| CN105527445B (en) * | 2015-12-30 | 2018-04-17 | 天津诺星生物医药科技有限公司 | A kind of acute myocardial infarction detecting system |
| CN107505459A (en) * | 2017-07-03 | 2017-12-22 | 广州瑞博奥生物科技有限公司 | Quantitatively detect people H FABP time-resolved fluoroimmunoassay chromatograph test strip, kit and preparation method thereof |
| CN107505459B (en) * | 2017-07-03 | 2019-12-24 | 广州瑞博奥生物科技有限公司 | Time-resolved fluorescence immunochromatographic test strip and kit for quantitatively detecting human H-FABP and preparation method thereof |
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