CN204514933U - B2M immunochromatographiassay assay quantitative detection test paper - Google Patents
B2M immunochromatographiassay assay quantitative detection test paper Download PDFInfo
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- CN204514933U CN204514933U CN201420648423.XU CN201420648423U CN204514933U CN 204514933 U CN204514933 U CN 204514933U CN 201420648423 U CN201420648423 U CN 201420648423U CN 204514933 U CN204514933 U CN 204514933U
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Abstract
The utility model discloses a kind of B2M 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 Streptavidin (or Avidin) mark fluorescent albumen and biotin labeled B2M monoclonal antibody; Described nitrocellulose membrane has detection line and nature controlling line, detection line is by B2M monoclonal antibody, and it has different identification epi-positions from above-mentioned biotin labeled B2M monoclonal antibody.The utility model is (as: chemoluminescence method/Immune-enhancing effect turbidimetry/colloidal gold method) compared with the method for detection B2M 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 B2M immunochromatographiassay assay quantitative detection test paper.
Background technology
B2M (β 2-microglobulin is called for short β 2-MG) is a kind of Small molecular globulin produced by lymphocyte, blood platelet, polymorphonuclear leukocyte, and molecular mass is 11800, the single chain polypeptide be made up of 99 amino acid.Extensively be present in blood plasma, urine, cerebrospinal fluid, saliva and first Ruzhong.The synthetic ratio of normal person's B2M and quite constant from the burst size cell membrane, B2M freely can filter from glomerulus, 99.9% absorbs at proximal tubular, be amino acid in local by metabolic degradation, so the discharge of B2M is very micro-under normal circumstances, be about 340mg from glomerular filtration every day, but in urine maximum excretion only have 370 μ g every day, only accounts for and filters 0.1% of total amount.
The rising of serum beta-2-microglobulin can reflect the situation whether impaired or filtered load of detection of glomeruli filtration function increases; And discharge B2M in urine and increase, then point out tubular injury or filtered load to increase; When urgent, chronic nephropyeltis, because of compromised kidneys, therefore Urine β2-microglobulin raises, and cystitis patients then B2M is normal; Renal transplant recipients blood, Urine β2-microglobulin obviously increase, and prompting body generation rejection, because B2M synthesis is accelerated, increase, and Serumβ2-microglobulins CD4+ cell counts still increases though kidney is removed.Therefore measure B2M in blood, urine to can be used for estimating hepatitis, ephritis, rheumatoid arthritis, and the active level of the disease such as malignant tumour, immunity disease, and as one that observes curative effect of medication easy, accurate and sensitive indicator.So being determined at of B2M has various value clinically.The common method of current detection B2M has:
1 particle reinforce immunoturbidimetry:
B2M in this method sample is formed immune complex with bag by the anti-human B2M on latex particle, produce turbidity and sample in B2M content proportional, measure by turbidimetry, thus try to achieve B2M content in sample.This method is easy, quick, accurate.
2 radio immunoassays:
Radiating immuning analysis technology utilizes isotope-labeled and unlabelled antigen, the method for synantibody generation Reverse transcriptase reaction.This method is a kind of highly sensitive, easier mensuration, but it needs to possess most advanced and sophisticated complicated equipment, and cost is not low yet.Meanwhile, this technology also needs special preventive measure, because it will use radiomaterial, there is the problem such as radiation and pollution.Therefore, nowadays radio immunoassay to a great extent replace by ELISA.
3 enzyme linked immunosorbent assay analysis methods:
This method B2M antibody bag is made insolubilized antibody by microwell plate, people's B2M is added in the micropore that Sheet is anti-, the B2M antibody marked with horseradish peroxidase (HRP) is again combined, form antibody-antigene-hrp-antibody complex, after thorough washing, add substrate tetramethyl benzidine (TMB) colour developing, the B2M in the depth of color and sample is proportionate.This method has fast, responsive, easy, be easy to the advantages such as standardization.
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, and (binding constant is up to 10 to have very high affinity to biology
15m
-1).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 B2M 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 B2M immunochromatographiassay assay quantitative detection test paper, it provides high sensitivity B2M 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: B2M 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 B2M monoclonal antibody and marked by streptavidin, the fixing nature controlling line identifying the detection line that the monoclonal antibody of B2M another one epi-position is formed and sheep anti-mouse igg polyclonal antibody formation on nitrocellulose filter in described detection zone.
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 rectangular opening.
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 the embodiment one shown in Fig. 1, B2-microglobulin 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, described fluorescent marker pad 2 is fixed with the fluorescin of biotin labeled B2M monoclonal antibody (concentration 0.3 ~ 1.5mg/mL) and marked by streptavidin (or Avidin), 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 B2M another one epi-position is formed, nature controlling line C is used for the validity of test strip.
The fluorescin that biotin labeled B2M 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 B2M 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, in sample, on B2M and fluorescent marker pad 2, the B2M antibody (Mab-β 2-MG*Fluoro) of combined with fluorescent label (fluorescin) reacts and forms compound β 2-MG-Mab-β 2-MG*Fluoro, compound is reacted when continuing to be advanced past B2M antibody (detection line) nitrocellulose membrane wrapping quilt under chromatography effect, B2-MG antibody capture formation compound (Mab-β 2-MG-β 2-MG-Mab-β 2-MG*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 B2M in sample, obtain B2M testing result.
As the further improvement of technique scheme, with reference to the embodiment three shown in 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 the embodiment two shown in Fig. 2 and Fig. 4 and embodiment four, 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 rectangular opening.
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. B2M immunochromatographiassay assay quantitative detection test paper, it is characterized in that: it comprises base plate, described base plate is provided with successively sample pad, 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 B2M monoclonal antibody and marked by streptavidin; The fixing nature controlling line identifying the detection line that the monoclonal antibody of B2M another one epi-position is formed and sheep anti-mouse igg polyclonal antibody formation on nitrocellulose filter in described detection zone.
2. B2M 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. B2M 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 B2M 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. B2M immunochromatographiassay assay quantitative detection test paper according to claim 4, is characterized in that: described view window is rectangular opening.
6. B2M immunochromatographiassay assay quantitative detection test paper according to claim 4, is characterized in that: described fluorescin is the one in green fluorescent protein, phycobniliprotein.
7. B2M 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. B2M immunochromatographiassay assay quantitative detection test paper according to claim 4, is characterized in that: described base plate is polystyrene component or tygon component.
9. B2M 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 |
|---|---|---|---|---|
| CN107132348A (en) * | 2017-02-22 | 2017-09-05 | 江苏雷森生物科技有限公司 | A kind of method of color fluorescence particle marker antibody and the test strips prepared using it |
| CN109575133A (en) * | 2018-12-28 | 2019-04-05 | 江苏众红生物工程创药研究院有限公司 | Anti-human β2- MG antibody and its application |
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2014
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107132348A (en) * | 2017-02-22 | 2017-09-05 | 江苏雷森生物科技有限公司 | A kind of method of color fluorescence particle marker antibody and the test strips prepared using it |
| CN109575133A (en) * | 2018-12-28 | 2019-04-05 | 江苏众红生物工程创药研究院有限公司 | Anti-human β2- MG antibody and its application |
| CN109575133B (en) * | 2018-12-28 | 2022-04-05 | 江苏众红生物工程创药研究院有限公司 | Anti-human beta2-MG antibodies and uses thereof |
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