CN208367017U - Serum amyloid A protein immunochromatographiassay assay quantitative detection test paper - Google Patents
Serum amyloid A protein immunochromatographiassay assay quantitative detection test paper Download PDFInfo
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- CN208367017U CN208367017U CN201820492324.5U CN201820492324U CN208367017U CN 208367017 U CN208367017 U CN 208367017U CN 201820492324 U CN201820492324 U CN 201820492324U CN 208367017 U CN208367017 U CN 208367017U
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Abstract
The utility model discloses a kind of serum amyloid A protein immunochromatographiassay assay quantitative detection test papers, including bottom plate, sample pad, fluorescent marker bonding pad, nitrocellulose and water absorption pad；The test strips are successively overlapped to be pasted on bottom plate and be constituted by sample pad, fluorescent marker bonding pad, nitrocellulose, water absorption pad.The serum amyloid A protein monoclonal antibody of biotin labeling and the fluorescin of marked by streptavidin are fixed on the fluorescent marker bonding pad；There are detection line and nature controlling line on the nitrocellulose membrane, detection line has different identification epitopes by serum amyloid A protein monoclonal antibody, from the serum amyloid A protein monoclonal antibody of above-mentioned biotin labeling.The utility model is compared with the method for detection serum amyloid A protein common at present, and such as: chemoluminescence method/Immune-enhancing effect turbidimetry/colloidal gold method not only substantially reduces detection time, while also improving detection sensitivity.
Description
Technical field
The utility model belongs to field of immunodetection, and in particular to a kind of immune layer of high sensitivity serum amyloid A
Analyse quantitative testing test paper item.
Background technique
Serum amyloid A protein (serumamyloidAprotein, SAA) is a kind of Acute reaction protein, is belonged to
Heterogeneous proteinoid in apolipoprotein family, relative molecular weight are about 12000.In acute-phase response, such as inflammation or sense
Contaminate acute stage, through interleukin 1 (interleukin-1, IL-1), interleukin-6 (interleukin-1, IL-6) and
Tumor necrosis factor a (tumornecrosisfactor-a, TNF-a) stimulates, and the macrophage and fiber being activated in liver are female
Cell can synthesize rapidly SAA, and concentration increases rapidly in 4-6h, be increased to 1000mg/L or more from the 1-5mg/L of normal level,
8-12h reaches peak value, but half life is short, only 50min or so, and declines rapidly in Recovery Phase of diseases.Serum amyloid A protein
Related with high-density lipoprotein (HDL), it can adjust the metabolism of high-density lipoprotein during inflammation.Certain diseases, such as virus
Infection, graft-rejection, dynamic atherosclerosis, the raising that SAA in blood can be detected in cardiovascular disease.
On the one hand, serum amyloid A protein is similar with C reactive protein (CRP), is reflection infectious diseases Earlier period of inflammation
Sensitive indicator, facilitate diagnose inflammation, assess its activity, monitor its activity and treatment.On the other hand, in certain diseases, SAA
Than other, common acute protein has more sensitivity, and such as in virus and bacterium infection, the serum-concentration of SAA is increased, and
In virus infection reaction, the serum-concentration of CRP hardly increases or increases unobvious.In infectious diseases, SAA it is absolute on
It rises and is higher than CRP, therefore SAA can provide better identification for small Acute-phase protein.For the trouble of kidney transplantation exclusion reaction
The diagnosis of person and the cystic fibrosis patient with adrenocortical hormones in treating, SAA ratio CRP are more acurrate.In addition to this, in class
In terms of rheumathritis, tuberculosis or leprosy, the chronic raising of SAA concentration is the prerequisite for synthesizing AA- starch fiber,
Also diagnosis secondary amyloidosis is used to become.Therefore the clinic that is measured as of SAA provides better diagnostic value.
At present detection serum amyloid A protein common method have enzyme linked immunosorbent assay (ELISA), immunoturbidimetry,
Colloidal gold immunity chromatography, fluorescence immune chromatography method.Wherein the ELISA method testing time is long, cumbersome, needs special instrument
Device is tested with the personnel Jing Guo professional training, while environmental limits is too many, is not suitable for clinical quick diagnosis.It is immune
Turbidimetry accuracy is lower, and anti-interference ability is weak, false positive and false negative result easily occurs, and antigen or antibody excess will lead to
Error is larger, and repeatability is bad.Colloidal gold method sensitivity is relatively poor, can not detect low content sample, commonly used in qualitative
Or sxemiquantitative test, do not meet clinical quantitative detection demand.Fluorescence immune chromatography method is swashed using fluorescent material in particular excitation light
The characteristic for generating fluorescence signal is given, has the advantages that high sensitivity, high specificity, reproducible, this method has been applied at present
In SAA clinical detection, and quickly grow.
Biotin-avidin system (biotinavidin system, BAS) is a kind of new bio reaction amplification system
System.With the appearance of various biotin derivatives, BAS is widely used in each field of medicine quickly.The system is applied to exempt from
Epidemic disease group, enzyme linked immunological, fluorescence immunoassay in the detection techniques such as radio-immunity, improve the sensitivity of the above technical method in which can dramatically
Property, specificity and stability, keep method easier, facilitate clinical quick diagnosis, and be conducive to carry out extensive epidemiology tune
It looks into, becomes the powerful of research immune response.It pollutes, is not required to multiple since BAS detection system economy is quick, and without radiogen
Miscellaneous instrument, a possibility that sufficiently showing the great potential and application of this system.
Biotin-avidin system testing principle: BAS is on the basis of conventional ELISA principle, in conjunction with biotin and parent
Highly enlarged effect between element, and a kind of detection system established.Avidin is a kind of alkalinity sugar extracted in ovalbumin
Albumen, molecular weight 68kDa are made of 4 subunits, and being known as very high affinity to biology, (binding constant is up to
1015M-1).Biotin is very easily with protein (such as antibody) with Covalent bonding together.In this way, combine the avidin molecule of enzyme with
The biotin molecule for being combined with specific antibody generates reaction, has not only played multistage amplification, but also since enzyme is encountering accordingly
Catalytic action and colour generation when substrate achieve the purpose that detect unknown antigen (or antibody) molecule.Streptavidin be with it is affine
It is known as a kind of protein of similar biological properties, every peptide chain can be in conjunction with one in molecule as Avidin for Streptavidin
A biotin, because nearly all substance for label can be combined with Avidin or streptavidin.Utilize biotin-parent
Detecting examination fast diagnosis reagent with plain system development serum amyloid A protein there is no report.
Utility model content
The purpose of this utility model is to provide a kind of serum amyloid A protein immunochromatographiassay assay quantitative detection test paper,
Based on double antibody sandwich method, fluorescence immunoassay lateral chromatography and biotin-Streptavidin amplification system, high sensitivity serum is provided
Amyloid A immunochromatographiassay assay quantitative detection test paper.When using the utility model, improves detection sensitivity and detection is stablized
Property, non-specific binding is reduced, detection time is not more than 10 minutes, substantially increases diagnosis efficiency.
The solution that the utility model solves its technical problem is: serum amyloid A protein immunochromatography quantitative detection
Test strips comprising bottom plate is successively arranged sample pad, fluorescent marker bonding pad, nitrocellulose filter and suction on the bottom plate
Water cushion, the water absorption pad and fluorescent marker bonding pad overlap be pressed in behind the both ends of nitrocellulose filter in nitrocellulose respectively
The surface of film forms detection zone, and the sample pad is overlapping to be pressed on fluorescent marker bonding pad, the fluorescent marker bonding pad
On be fixed with the serum amyloid A protein monoclonal antibody of biotin labeling and the fluorescin of marked by streptavidin；It is described
The monoclonal antibody of fixed identification another epitope of serum amyloid A protein is constituted on nitrocellulose filter in detection zone
The nature controlling line that detection line and sheep anti-mouse igg polyclonal antibody are constituted.
As a further improvement of the above technical scheme, the fluorescent marker bonding pad includes the first fluorescence mark of stacking
Remember object bonding pad and the second fluorescent marker bonding pad, one end of the first fluorescent marker bonding pad is padded under sample pad
Side, the overlapping one end for being pressed in nitrocellulose filter of the second fluorescent marker bonding pad.
As a further improvement of the above technical scheme, the detection line is located in detection zone combines close to fluorescent marker
One end of pad, the nature controlling line are located at one end in detection zone close to water absorption pad, and detection line and nature controlling line are separated by a distance for 4-
8mm。
It as a further improvement of the above technical scheme, further include getting stuck, the bottom plate, sample pad, fluorescent marker knot
Close pad, in nitrocellulose filter and water absorption pad are placed in and get stuck, the position in the shell surface that gets stuck corresponding to nitrocellulose membrane is set
There is observation window, is equipped with well corresponding to the position of sample pad in the shell surface that gets stuck.
As a further improvement of the above technical scheme, the observation window is slot.
As a further improvement of the above technical scheme, the fluorescin is green fluorescent protein, in phycobniliprotein
It is a kind of.
As a further improvement of the above technical scheme, the sample pad is after surfactant buffer immersion treatment
Dry glass fibre component.
As a further improvement of the above technical scheme, the bottom plate is polystyrene component or polyethylene component.
As a further improvement of the above technical scheme, described to get stuck including plastics upper casing and plastics lower casing, the plastics
Upper casing is formed after fastening on plastics lower casing to get stuck.
The utility model compared with prior art, has the advantages that
(1) using fluorescin as marker, this marker has good stability the utility model, is conducive to improve detection surely
It is qualitative.
(2) the utility model improves detection sensitivity, reduces by utilizing " Streptavidin-biotin amplification system "
Non-specific binding is conducive to improve kit performance.
(3) it is not more than 10 minutes using the detection time that the utility model carries out serum amyloid A protein, detects linear model
It encloses for 3.0mg/L~200.00mg/L, greatly improves detection efficiency.
(4) the utility model can carry out interpretation to result by fluorescence immunity analyzer, it can be achieved that automation, is reduced subjective
The influence of factor provides convenient, quick, reliable diagnostic result.
(5) well entered equipped with sample and for the observation window of observed result of getting stuck in the utility model, according to point
Analyzer device determines as a result, accurate and reliable.
(6) the utility model is easy to make, small in size, easy to carry.
(7) the utility model testing cost is lower.
(8) the utility model can be mass, and be suitable for clinical quick diagnosis and live quick diagnosis；It is easy to save, has
Conducive to grass-roots unit's popularization.
Detailed description of the invention
It, below will be to required in embodiment description in order to illustrate more clearly of the technical scheme in the embodiment of the utility model
Attached drawing to be used is briefly described.Obviously, described attached drawing is a part of the embodiment of the utility model, rather than complete
Portion's embodiment, those skilled in the art without creative efforts, can also be obtained according to these attached drawings it
His design scheme and attached drawing.
Fig. 1 is the structural schematic diagram of the utility model embodiment one；
Fig. 2 is the structural schematic diagram of the utility model embodiment two；
Fig. 3 is the structural schematic diagram of the utility model embodiment three；
Fig. 4 is the structural schematic diagram of the utility model embodiment four.
Specific embodiment
It is carried out below with reference to technical effect of the embodiment and attached drawing to the design of the utility model, specific structure and generation
It clearly and completely describes, to be completely understood by the purpose of this utility model, feature and effect.Obviously, described embodiment
It is a part of the embodiment of the utility model, rather than whole embodiments, it is based on the embodiments of the present invention, the skill of this field
Art personnel other embodiments obtained without creative efforts belong to the model of the utility model protection
It encloses.In addition, all connection/connection relationships being previously mentioned in text, not singly refer to that component directly connects, and referring to can be according to specific reality
Situation is applied, by adding or reducing couple auxiliary, to form more preferably coupling structure.
Referring to Fig.1, serum amyloid A protein immunochromatographiassay assay quantitative detection test paper comprising bottom plate 5, on the bottom plate 5
It is successively arranged sample pad 1, fluorescent marker bonding pad 2, nitrocellulose filter 3 and water absorption pad 4, the water absorption pad 4 and fluorescence mark
Overlapping be pressed in behind the both ends of nitrocellulose filter 3 forms detection zone on the surface of nitrocellulose filter 3 to note object bonding pad 2 respectively,
Fluorescent marker bonding pad 2 is glass fibre membrane, and the sample pad 1 is overlapping to be pressed on fluorescent marker bonding pad 2, the fluorescence
The serum amyloid A protein monoclonal antibody (0.3~1.5mg/mL of concentration) of biotin labeling is fixed on marker bonding pad 2
(excitation wavelength 530nm, wavelength of transmitted light 570nm, concentration are used with the fluorescin of marked by streptavidin (or Avidin)
0.1~1.0mg/mL)；Be fixed on nitrocellulose filter 3 in the detection zone identification serum amyloid A protein another
The Quality Control that the detection line T (0.5~3mg/mL of concentration) and sheep anti-mouse igg polyclonal antibody that the monoclonal antibody of epitope is constituted are constituted
Line C (0.2~2.0mg/mL of concentration), nature controlling line C are used for the validity of test strip.
The serum amyloid A protein monoclonal antibody and Streptavidin (or Avidin) of biotin labeling are marked glimmering
Photoprotein is fixed on fluorescent marker bonding pad 2, will have the monoclonal of identification another epitope of serum amyloid A protein to resist
Body and sheep anti-mouse igg polyclonal antibody are fixed on nitrocellulose filter respectively as detection line T and nature controlling line C.When to test sample
It after product are added in sample pad 1, are acted on and being moved forward by chromatography, serum amyloid A protein is in conjunction with fluorescent marker in sample
Serum amyloid A protein antibody (Mab-SAA*Fluoro) reaction on pad 2 in conjunction with fluorescent marker (fluorescin) forms multiple
Object SAA-Mab-SAA*Fluoro is closed, reaction compound is continued to move along by wrapping on nitrocellulose membrane under chromatography effect
When SAA antibody (detection line) of quilt, the coated SAA antibody capture of reaction compound forms compound (Mab-SAA-SAA-
Mab-SAA*Fluoro) (detection line) reads the reaction signal of detection line by fluorescence immunity analyzer, in the work of excitation light source
Under, fluorescent material emit specific wavelength fluorescence signal, fluorescence immunity analyzer capture fluorescence signal, by signal conversion and
The standard curve of setting is automatically converted to quantitative value, calculates the concentration of serum amyloid A protein in sample, obtains serum shallow lake
Powder sample albumin A testing result.
As a further improvement of the above technical scheme, referring to Fig. 3, the fluorescent marker bonding pad 2 includes stacking
First fluorescent marker bonding pad 20 and the second fluorescent marker bonding pad 21, the one of the first fluorescent marker bonding pad 20
End pad is in the lower section of sample pad 1, the overlapping one end for being pressed in nitrocellulose filter 3 of the second fluorescent marker bonding pad 21.It is glimmering
The layering of signal object bonding pad 2 is arranged, convenient for fluorescent marker bonding pad 2 is mounted on sample pad 1 and nitrocellulose filter 3
Between.
As a further improvement of the above technical scheme, the detection line T referring to described in Fig. 3 is located in detection zone close to fluorescence mark
Remember one end of object bonding pad 2, the nature controlling line C is located at one end in detection zone close to water absorption pad 4, detection line T and nature controlling line C phase
Every distance be 4-8mm.
It as a further improvement of the above technical scheme, further include getting stuck referring to Fig. 2 and Fig. 4, the bottom plate 5, sample pad
1, fluorescent marker bonding pad 2, nitrocellulose filter 3 and water absorption pad 4, which are placed in, gets stuck in 6, corresponds in 6 shell surfaces that get stuck
The position of nitrocellulose membrane 3 is equipped with observation window 8, and the position that sample pad 1 is corresponded in 6 shell surfaces that get stuck is equipped with well 7.
As a further improvement of the above technical scheme, the observation window 8 is slot.
As a further improvement of the above technical scheme, the fluorescin is green fluorescent protein, in phycobniliprotein
It is a kind of.
As a further improvement of the above technical scheme, the sample pad 1 is through surfactant buffer immersion treatment
Dry glass fibre component afterwards.Sample pad is made of glass fibre, through surfactant buffer immersion treatment, is made after dry
With.The cutting width of the sample pad 1 is 0.3~0.5cm.
As a further improvement of the above technical scheme, the bottom plate 5 is polystyrene component or polyethylene component.
As a further improvement of the above technical scheme, it is described get stuck 6 include plastics upper casing 60 and plastics lower casing 61, it is described
Plastics upper casing 60 is formed after fastening on plastics lower casing 61 gets stuck 6.
It is to be illustrated to the better embodiment of the utility model, but the invention is not limited to institute above
Embodiment is stated, those skilled in the art can also make various be equal without departing from the spirit of the present invention
Variation or replacement, these equivalent variation or replacement are all included in the scope defined by the claims of the present application.
Claims (9)
1. serum amyloid A protein immunochromatographiassay assay quantitative detection test paper, it is characterised in that: it includes bottom plate, on the bottom plate
It is successively arranged sample pad, fluorescent marker bonding pad, nitrocellulose filter and water absorption pad, the water absorption pad and fluorescent marker knot
Closing pad, overlapping be pressed in behind the both ends of nitrocellulose filter forms detection zone, the sample pad on the surface of nitrocellulose filter respectively
It is overlapping to be pressed on fluorescent marker bonding pad, the serum amyloid sample of biotin labeling is fixed on the fluorescent marker bonding pad
The fluorescin of albumin A monoclonal antibody and marked by streptavidin；It is fixed on nitrocellulose filter in the detection zone to know
The detection line and sheep anti-mouse igg polyclonal antibody structure that the monoclonal antibody of another epitope of other serum amyloid A protein is constituted
At nature controlling line.
2. serum amyloid A protein immunochromatographiassay assay quantitative detection test paper according to claim 1, it is characterised in that: institute
State the first fluorescent marker bonding pad and the second fluorescent marker bonding pad that fluorescent marker bonding pad includes stacking, described the
For one end pad of one fluorescent marker bonding pad in the lower section of sample pad, the second fluorescent marker bonding pad is overlapping to be pressed in nitric acid
One end of cellulose membrane.
3. serum amyloid A protein immunochromatographiassay assay quantitative detection test paper according to claim 2, it is characterised in that: institute
One end that detection line is located at close fluorescent marker bonding pad in detection zone is stated, the nature controlling line is located in detection zone close to water suction
One end of pad, detection line and nature controlling line are separated by a distance for 4-8mm.
4. serum amyloid A protein immunochromatographiassay assay quantitative detection test paper according to any one of claims 1 to 3, special
Sign is: further including getting stuck, the bottom plate, sample pad, fluorescent marker bonding pad, nitrocellulose filter and water absorption pad are placed in
In getting stuck, it is equipped with observation window corresponding to the position of nitrocellulose membrane in the shell surface that gets stuck, corresponds to sample pad in the shell surface that gets stuck
Position be equipped with well.
5. serum amyloid A protein immunochromatographiassay assay quantitative detection test paper according to claim 4, it is characterised in that: institute
Stating observation window is slot.
6. serum amyloid A protein immunochromatographiassay assay quantitative detection test paper according to claim 4, it is characterised in that: institute
Stating fluorescin is one of green fluorescent protein, phycobniliprotein.
7. serum amyloid A protein immunochromatographiassay assay quantitative detection test paper according to claim 4, it is characterised in that: institute
Stating sample pad is dry glass fibre component after surfactant buffer immersion treatment.
8. serum amyloid A protein immunochromatographiassay assay quantitative detection test paper according to claim 4, it is characterised in that: institute
Stating bottom plate is polystyrene component or polyethylene component.
9. serum amyloid A protein immunochromatographiassay assay quantitative detection test paper according to claim 4, it is characterised in that: institute
It states and gets stuck including plastics upper casing and plastics lower casing, the plastics upper casing is formed after fastening on plastics lower casing to get stuck.
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| CN112924686A (en) * | 2019-12-05 | 2021-06-08 | 复旦大学 | Immunochromatography test strip for detecting serum amyloid A and preparation and detection methods thereof |
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| CN110488006A (en) * | 2019-09-26 | 2019-11-22 | 天津华科泰生物技术有限公司 | A kind of immunochromatographydetection detection card and preparation method thereof of 3 sample albumen 1 of quick detection chitinase |
| CN111656197A (en) * | 2019-10-15 | 2020-09-11 | 湖南乾康科技有限公司 | Test strip and method for detecting urine amyloid A beta protein |
| WO2021072651A1 (en) * | 2019-10-15 | 2021-04-22 | 湖南乾康科技有限公司 | TEST STRIP AND METHOD FOR MEASURING URINE Aβ AMYLOID PROTEIN |
| CN111656197B (en) * | 2019-10-15 | 2021-10-19 | 湖南乾康科技有限公司 | Test strip and method for detecting urine amyloid A beta protein |
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| JP2022539580A (en) * | 2019-10-15 | 2022-09-12 | 湖南乾康科技有限公司 | Test strips and methods for detecting urinary Aβ amyloid |
| JP7385944B2 (en) | 2019-10-15 | 2023-11-24 | 湖南乾康科技有限公司 | Test strip and method for detecting Aβ amyloid in urine |
| CN112924686A (en) * | 2019-12-05 | 2021-06-08 | 复旦大学 | Immunochromatography test strip for detecting serum amyloid A and preparation and detection methods thereof |
| WO2024087430A1 (en) * | 2023-02-28 | 2024-05-02 | 湖南乾康科技有限公司 | Enzyme-linked immunosorbent assay method for quantitatively detecting urine amyloid precursor protein fragment |
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