CN204514932U - Myoglobins immunochromatographiassay assay quantitative detection test paper - Google Patents
Myoglobins immunochromatographiassay assay quantitative detection test paper Download PDFInfo
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- CN204514932U CN204514932U CN201420648353.8U CN201420648353U CN204514932U CN 204514932 U CN204514932 U CN 204514932U CN 201420648353 U CN201420648353 U CN 201420648353U CN 204514932 U CN204514932 U CN 204514932U
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Abstract
The utility model discloses a kind of myoglobins 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 myoglobins monoclonal antibody; Described nitrocellulose membrane has detection line and nature controlling line, detection line is by myoglobins monoclonal antibody, and it has different identification epi-positions from above-mentioned biotin labeled myoglobins monoclonal antibody.The utility model is (as: chemoluminescence method/Immune-enhancing effect turbidimetry/colloidal gold method) compared with the method for detection myoglobins 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 myoglobins immunochromatographiassay assay quantitative detection test paper.
Background technology
Myoglobins (Myoglobin, MYO), the associated proteins be made up of a peptide chain and a prosthetic heme group is the protein storing oxygen in muscle, is also the main protein of composition skeletal muscle and cardiac muscle.When muscle damage, MYO can drain in blood circulation from musculature, and serum MYO concentration is increased, and this index is used for judging whether muscle damage occurs.Measure serum MYO, can be used as the sensitiveest early stage index that acute myocardial infarction AMI (AMI) is diagnosed, also can be used as an index of observing muscle damage situation at surgery operative connection.
At present, detect equal Shortcomings in all multi-methods of myocardial injury markers, as: euzymelinked immunosorbent assay (ELISA), solid-phase immunity layer inhale method, radio immunoassay, complex operation, detection time length, the pollution of result poor repeatability, nucleic, some method can only carry out qualitative detection.The advantage that though chemiluminescence immunoassay has is accurate, highly sensitive, high specificity, precision are good, its instrument is expensive, and agents useful for same is again import reagent, and common laboratory is difficult to carry out, and is not suitable for emergency treatment inspection.
The common method of current detection MYO has:
1 Latex Agglutination:
Latex Agglutination is clinical comparatively conventional serological method, the anti-human MYO antibody sensitized of emulsion reagent purifying, can and patients serum in MYO generation specific reaction, present agglutinating particle clearly in several minutes, occur that aggegation person is positive, do not occur that aggegation person is for negative.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.
2 Immunity transmission turbidity:
Immunity transmission turbidity principle is, utilize antigen and antibody specific binding formed compound, by measure complex formation number quantitative method is carried out to antigen or antibody.This method is easy and simple to handle, and be applicable to common automatic biochemistry analyzer and common spectrophotometer, nearly all laboratory all can be carried out.Unfortunately sensitivity and precision are all not ideal enough, and required antiserum amount is large, and the cycle of detection is longer.
3 radioimmunologies:
Radioimmunology is the sensitiveest serum MYO assay method, utilize MYO to mark MYO with 125I and compete limited antibody, minimum measurement range is the level of 2ng/ml, due to radioimmunology, to there is the radioactive isotope half life period short, the shortcomings such as radioactive contamination is not easily preserved, poor stability, have inconvenience in using, especially the widespread use of enzyme immunoassay, makes the method seldom adopt clinically now.
Biotin-avidin system (biotinavidin system, BAS), is a kind of new bio reaction amplification system, is widely used in each field of medical science.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.
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 myoglobins 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 myoglobins immunochromatographiassay assay quantitative detection test paper, it provides high sensitivity myoglobins 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: myoglobins 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 myoglobins 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 myoglobins 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 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, myoglobins 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 myoglobins 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 myoglobins another one epi-position is formed, nature controlling line C is used for the validity of test strip.
The fluorescin that biotin labeled myoglobins 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 myoglobins 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 myoglobins and fluorescent marker pad 2, the myoglobins antibody (Mab-MYO*Fluoro) of combined with fluorescent label (fluorescin) reacts and forms compound MYO-Mab-MYO*Fluoro, compound is reacted when continuing to be advanced past MYO antibody (detection line) nitrocellulose membrane wrapping quilt under chromatography effect, MYO antibody capture formation compound (Mab-MYO-MYO-Mab-MYO*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 myoglobins in sample, obtain myoglobins 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. myoglobins 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 myoglobins 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 myoglobins another one epi-position is formed.
2. myoglobins 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. myoglobins 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 myoglobins 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. myoglobins immunochromatographiassay assay quantitative detection test paper according to claim 4, is characterized in that: described view window is rectangular opening.
6. myoglobins 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. myoglobins 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. myoglobins immunochromatographiassay assay quantitative detection test paper according to claim 4, is characterized in that: described base plate is polystyrene component or tygon component.
9. myoglobins 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 (1)
| 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 |
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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 |
| 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 |
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