CN104849264A - Chemiluminiscence immune test strip and preparation method thereof - Google Patents
Chemiluminiscence immune test strip and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a chemiluminiscence immune test strip and a preparation method thereof. The immune test strip comprises a sample dropping region, an antigen capturing region and a chemiluminiscence detection region, wherein the sample dropping region is prepared from a high-hydrophily material and is used for dropping a sample to be detected; an antibody, an antigen and/or an antiantibody which are used for capturing molecules to be detected in the sample to be detected are fixed in the antigen capturing region; transparent micro particles are fixed in the chemiluminiscence detection region; an antibody, an antigen and/or an antiantibody which can be bonded with the molecules to be detected in the sample to be detected as well as a chemiluminiscence substrate, a stabilizer and/or a stabilizer are modified on the surfaces of the transparent micro particles. Under the combination amplification action of the transparent micro particles, weak light sensitivity detection of the sample to be detected is realized; the transparent micro particles are prepared from a transparent organic macromolecular material. The chemiluminiscence immune test strip has the advantages of convenience in detection, high sensitivity, one-step quantitative detection, easiness in storage and the like; an external reagent is not needed, and quick and quantitative detection of a drop of blood sample or urine sample can be realized.
Description
Technical field
The present invention relates to immunity biosensor, chemiluminescence optical detection preparing technical field, relate more specifically to a kind of chemiluminescence immunoassay examination bar and preparation method thereof, this examination bar can detect protein macromolecule or other antigen molecule concentration in biological sample fast.
Background technology
For in the quick detection of the protein macromolecule in the biological sample such as blood sample, urine sample, immune detection is the most frequently used a kind of technology.Method is easy, specificity is high.Utilize antigen and antibody specific binding and the high-selectivity analysis method set up, further probe technique is introduced in analyzing to realize on-line checkingi.Immune detection comprises enzyme-linked immunosorbent assay, fluorescent immune method, electro-chemistry immunity method, Electrochemiluminescence immunoassay, chemiluminescence immunoassay etc.Immune detection not only can detect large molecule, as the various protein etc. in body fluid, can also detect some Small molecular, as the hormone, microbiotic, medicine etc. of small-molecular-weight.
POCT (point-of-care testing) is also referred to as real-time test, and POCT experimental technique instrument has the feature of miniaturization, simple operation, report the test just-in-time, has started new test mode, and patient can be made to obtain diagnostic result as early as possible.POCT experimental technique shortens checks turnover period, eliminates preprocessor and the time of sample complexity, can analyze in sampling location.At present, POCT products application increasing extent is extensive, it has actual clinical practice at comprehensive clinical labororatory, ward, Emergency call, operating room, ICU, ambulance, recovery centre, blood bank, family care mechanism, community, Rural medical clinic etc., and effect is better.
Dry chemical technology, that one or more reaction reagent dryings are fixed on solid carrier, such as, on the scraps of paper, film etc., use the liquid existed in sample as reaction medium, tested composition directly carries out colour generation, electric current or luminescence-producing reaction with the dry reagent solidified on carrier.
Along with the development of technology, there is cross development in immunological technique, POCT technology and dry chemical technology, and be born the such as technology such as colloid gold immune test paper detection and fluorescence immunoassay detection paper thus.But the detection sensitivity of colloid gold test paper is inadequate, quantitative color detects inaccurate, and usually can only be used for qualitative or semi-quantitative analysis, fluorescence immunoassay test paper needs special fluorescence detection device, and cost compare is high.
Summary of the invention
In view of this, the present invention is directed to the quick detection demand of the extremely low concentration materials such as protein macromolecule in sample, fundamental purpose is to provide a kind of chemiluminescence immunoassay to try bar and preparation method thereof, thus can realize simple and quick, highly sensitive quantitative detection.
To achieve these goals, as one aspect of the present invention, the invention provides a kind of chemiluminescence immunoassay examination bar, comprising:
Sample drips district, adopts hydrophilic material manufacture, and testing sample drips in this region;
Trapping region, is fixed with antibody, antigen and/or the antiantibody that can catch molecules detected in testing sample in described trapping region;
Chemiluminescence detection district, transparent particle is fixed with in described detection zone, described transparent particle sub-surface is modified with can molecules detected is combined in described testing sample antibody, antigen and/or antiantibody, and chemical luminous substrate, chemiluminescence stabilizing agent and/or reinforcing agent, due to transparent particle in conjunction with amplification, realize the Sensitive Detection of the faint light of testing sample;
Wherein, described transparent particle is prepared by transparent organic polymer material.
Wherein, the homogeneous transparent spheroid that described transparent particle is diameter between 100nm ~ 50 μm, described transparent particle be selected from polyvinyl alcohol microparticles, polyvinyl pyrrolidone microballoon, chitosan microball, PVA-PS complex microsphere, polyvinyl alcohol-gelatine microballoon, polypropylene microballoon, polyethyleneglycol diacrylate microballoon, PEG-DA/HEMA gel copolymer microballoon one of at least; And
Described transparent particle sub-surface has been carried out carboxyl, hydroxyl or sulfydryl hydrophile function group and has been modified.
Wherein, described chemical luminous substrate is luminol and derivant, different luminol and derivant thereof, 1,2-dichloroethane luminophor or dioxetane.
Wherein, when described chemiluminescence immunoassay examination strip adoption HRP luminophor, described chemiluminescence stabilizing agent and reinforcing agent select fluorescein, thiazole, iodophenol, 6-hydroxybenzothiazole, 4-iodobenzene boric acid and/or bromjophenol blue; When described chemiluminescence immunoassay examination strip adoption ALP luminescence system, described chemiluminescence stabilizing agent and reinforcing agent select polychlorostyrene benzyl ethylene benzyl Dimethyl Ammonium BDMQ, TMQ and/or BSA.
Wherein, when described transparent particle sub-surface is combined with antiantibody, described antiantibody has carried out biotin labeling.
Wherein, described trapping region comprises enzyme mark monoclonal antibody 1 land and monoclonal antibody 2 trapping region, described enzyme mark monoclonal antibody 1 land is fixed with enzyme mark-monoclonal antibody 1, and the antigen in testing sample is combined with described enzyme mark-monoclonal antibody 1, forms " enzyme mark-antibody 1-antigen " compound; Described monoclonal antibody 2 trapping region is fixed with monoclonal antibody 2, and the antigen in testing sample is combined with described monoclonal antibody 2, forms " enzyme mark-antibody 1-Ag-Ab 2 " compound; And
The unnecessary enzyme mark-monoclonal antibody 1 in described enzyme mark monoclonal antibody 1 land to be reacted and luminous along with the luminous substrate of transparent particle sub-surface in the flowing of liquid and described chemiluminescence detection district, thus in described testing sample, the concentration of antigen and the luminous intensity in described chemiluminescence detection district are negative correlation.
Wherein, the enzyme mark thing in described enzyme mark monoclonal antibody 1 land is horseradish peroxidase HRP or alkaline phosphatase.
As another aspect of the present invention, present invention also offers the preparation method of a kind of chemiluminescence immunoassay examination bar, comprise the following steps:
Step 1, on a transparent substrate, pre-bonding one deck water wettability thin paper, is stained with glass fibre element film, immobilized monoclonal antibody 2 on described glass fibre element film, dry;
Step 2, on nitrocellulose filter, the fixing transparent particle modified is sub; Described transparent particle is prepared by transparent organic polymer material, be selected from polyvinyl alcohol microparticles, polyvinyl pyrrolidone microballoon, chitosan microball, PVA-PS complex microsphere, polyvinyl alcohol-gelatine microballoon, polypropylene microballoon, polyethyleneglycol diacrylate microballoon, PEG-DA/HEMA gel copolymer microballoon one of at least, diameter is between 100nm ~ 50 μm, and described transparent particle sub-surface has been carried out carboxyl, hydroxyl or sulfydryl hydrophile function group and modified;
The glass fibre element paper of step 3, immobilized enzyme mark monoclonal antibody land, spraying monoclonal antibody linked with peroxidase 1, dry;
Step 4, fixed sample pad glass fibre element paper.
Step 5, slitting, hermetically drying is preserved.
Wherein, in step 2, fix described transparent particle by carving rectangle white space on described nitrocellulose filter, or fix described transparent particle by described transparent particle being painted on described nitrocellulose filter surface.
Wherein, the enzyme mark thing in the mark of enzyme described in step 3 monoclonal antibody 1 land is alkaline phosphatase, and described alkaline phosphatase has carried out marked by streptavidin.
Known based on technique scheme, chemiluminescence immunoassay examination bar of the present invention is higher relative to existing colloid gold immune examination bar detection sensitivity, can be combined, realize simple and quick, high-sensitive accurate quantification and detect with portable optical detector; Chemiluminescence immunoassay examination bar of the present invention combines chemiluminescence, immunological technique, chromatographic technique, particulate technology, achieve the fast quantification of material and the Site Detections of single step such as low concentration protein macromolecules in sample, can be used for emergency treatment detection, on-site quick screening or family and test oneself fast.
Accompanying drawing explanation
Fig. 1 is the structural representation of chemiluminescence immunoassay of the present invention examination bar;
Fig. 2 is the reaction schematic diagram of transparent particle of the present invention in luminous detection zone.
Embodiment
For making the object, technical solutions and advantages of the present invention clearly understand, below in conjunction with specific embodiment, and with reference to accompanying drawing, the present invention is described in further detail.
Colloid gold test paper detection technique insufficient sensitivity is high, generally can only be used for qualitative or half-quantitative detection; And existing chemiluminescence all needs many more manipulations usually, need the steps such as separation, be difficult to quick, the ease of Use that realize single step.The present invention is by combining immunological technique and chemiluminescence, combine the high selectivity of chemiluminescent high sensitivity and immuno analytical method, after the enzyme mark material that antibody marks is combined with luminous substrate, react generation photon, the intensity of photon and the concentration of determinand is proportionate or negative correlation, thus optics can be carried out by portable detector and quantitatively detect, draw the concentration of determinand quickly and easily.
In addition, usually in liquid chemiluminescence detecting step, determinand is combined the compound forming enzyme labelled antibody 1-Ag-Ab 2-magnetic bead with enzyme labelled antibody 1; After Magneto separate, Excess antibody is removed; Add luminous substrate, under enzyme effect, produce chemiluminescence.And the present invention is in order to reduce separation, washing, add the reactions steps such as luminous substrate, and combine with chromatography strip technology, the reaction trying bar from general liquid phase immunochemiluminescence and chromatography is all different, formation enzyme labelled antibody 1-Ag-Ab 2 compound is fixed on to catch to be brought, unnecessary enzyme labelled antibody 1 reacts with luminous substrate, and antigen concentration and light intensity present the technical scheme of negative correlation.
Thus, the invention discloses a kind of chemiluminescence immunoassay examination bar, comprise sample and drip district, trapping region, chemiluminescence detection district, waste liquid suction zones and substrate.This chemiluminescence immunoassay examination bar may be used for the quick detection of the extremely low concentration materials such as protein macromolecule in biological sample.Described chemiluminescence immunoassay examination bar is the integrating test bar prepared based on chromatographic theory and immunochemiluminescence principle, wherein,
Sample drips district, and adopt high-hydrophilic material, namely testing sample drips in this region;
Trapping region, is fixed with antibody, antigen and/or the antiantibody that can catch molecules detected in testing sample in described trapping region;
As one, preferably trapping region comprises enzyme mark monoclonal antibody 1 land and antigen capture band, wherein,
Enzyme mark monoclonal antibody 1 land, be the storage area of enzyme mark-monoclonal antibody 1, the antigen in testing sample is combined with this enzyme mark-monoclonal monoclonal antibody 1, forms " enzyme mark-antibody 1-antigen " compound;
Antigen capture band, is also referred to as fixing monoclonal antibody 2 reaction zone, is the fixed area of monoclonal antibody 2, compound flow to this district, antigen is combined with immobilized monoclonal antibody 2, forms " enzyme mark-antibody 1-Ag-Ab 2 " compound, thus makes that this compound is captured is fixed on this region;
Chemiluminescence detection district, is prepared by transparent organic polymer material, is the fixed area of transparent particle in conjunction with antiantibody and luminous substrate, is fixed with chemiluminescence stabilizing agent and/or reinforcing agent simultaneously.Transparent particle sub-surface has modified two anti-(antiantibody) and efficient chemical luminous substrate, when excessive enzyme labelled antibody 1 flow to the region that this is fixed with transparent particle, two anti-specific recognition enzyme mark monoclonal antibodies 1, enzyme mark thing is combined with luminous substrate and stabilized illumination, and due to transparent particle in conjunction with amplification, achieve the Sensitive Detection of faint light, thus quantitatively can obtain the concentration of antigen.
Wherein, described sample drips district and adopts high porosint, and testing sample drips can be infiltrated sample pad rapidly in this region, and can movement fast, does not stay in sample pad storage; Sample pad material, such as, can adopt all-glass paper, filter paper etc.; Sample drips district to be needed to carry out pretreatment, and pretreatment fluid comprises large molecule water wetted material, protein protection material or surfactant.
In described enzyme mark monoclonal antibody 1 land, enzyme mark thing can be horseradish peroxidase (horseradish peroxidase, HRP) or alkaline phosphatase (alkaline phosphatase, ALP).Alkaline phosphatase can carry out marked by streptavidin.
Described enzyme mark monoclonal antibody 1 land can adopt preparation such as material such as glass fibre element paper, filter paper etc., and land material first carries out pre-service and after drying, sprays enzyme mark-monoclonal antibody 1, low temperature drying.
Described antigen capture band adopts the preparation of cellulose nitrate film material, for the fixed area of monoclonal antibody 2, monoclonal antibody 2 adopts spraying or hatched manner to be fixed on cellulose nitrate membrane material, when compound flow to this district, antigen in compound is combined with fixing monoclonal antibody 2, form " enzyme mark-antibody 1-Ag-Ab 2 " compound, thus make that this compound is captured is fixed on this region.
In the sub-chemiluminescence detection band of described transparent particle, the homogeneous transparent spheroid that transparent particle is diameter between 100nm ~ 50 μm, hydrophilicity-imparting treatment has been carried out on surface, such as adopt carboxyl, the hydrophile function such as hydroxyl or sulfydryl group modifies, transparent particle can be such as ST-EVB (styrene and divinylbenzene copolymer), PMMA (polymethacrylate), PVA (hydrophilic polyethene alcohol), PVP (polyvinyl pyrrolidone), shitosan (CS), PVA-PS complex microsphere, PET (polyethylene terephthalate) microballoon, polyvinyl alcohol-gelatine, PAA (polypropylene), polyethyleneglycol diacrylate (PEGDA), one or more in the materials such as PEG-DA/HEMA gel copolymer microballoon.
In the sub-chemiluminescence detection band of described transparent particle, luminous substrate can be luminol, different luminol and derivant thereof, 1,2-dichloroethane luminophor, dioxetane etc.Stable luminescence agent and reinforcing agent, such as fluorescein, thiazole, iodophenol, 6-hydroxybenzothiazole, 4-iodobenzene boric acid, bromjophenol blue etc. can be used for HRP luminescence system; Or such as polychlorostyrene benzyl ethylene benzyl Dimethyl Ammonium (BDMQ), anti-aging agent RD (TMQ) or bovine serum albumin(BSA) (BSA) etc. can be used for ALP luminescence system.
The antiantibody that transparent particle sub-surface is modified can carry out biotin labeling.
Described waste liquid suction zones is prepared from by the material with strong absorptive, and unnecessary sample liquids is stored in herein by absorption.
Described substrate is the supporting construction of this chemiluminescence immunoassay examination bar, opaque propping material or transparent plastic substrate can be selected to make, thus in conjunction with different optical gauges, the optics being beneficial to distinct device quantitatively detects.Preferred substrate adopts nitrocellulose filter (NC film) manufacture.
The invention also discloses the preparation method of a kind of chemiluminescence immunoassay examination bar, comprise the following steps:
Step 1, on a transparent substrate, pre-bonding one deck water wettability thin paper, is stained with glass fibre element film, and immobilized monoclonal antibody 2 on glass fibre element film is dry; Glue thieving paper material.
Step 2, on nitrocellulose filter, the fixing transparent particle modified is sub; Rectangle white space can be carved thereon and fix this transparent particle, also this transparent particle can be painted on this nitrocellulose filter surface.
The glass fibre element paper of step 3, immobilized enzyme mark monoclonal antibody land, spraying monoclonal antibody linked with peroxidase 1, dry;
Step 4, fixed sample pad glass fibre element paper.
Step 5, slitting, hermetically drying is preserved.
Wherein, when adopting the fixing transparent particle period of the day from 11 p.m. to 1 a.m modified in the rectangular recess or fracture carved on nitrocellulose filter, transparent particle needs to modify.Transparent particle had both considered its transparency, considered the reagent sensitivity after its dried stability and redissolution and transparency again.The sub-microsphere surface of transparent particle combines antiantibody and luminous substrate, is fixed with chemiluminescence stabilizing agent and reinforcing agent simultaneously.
Below in conjunction with drawings and the specific embodiments, explanation is further elaborated to the solution of the present invention.
Fig. 1 shows the structural representation of the chemiluminescence immunoassay examination bar as a preferred embodiment of the invention.As shown in the figure, chemiluminescence immunoassay examination bar of the present invention comprises sample dropping district 1, enzyme mark monoclonal antibody 1 reaction zone 2, antigen capture band 3, the sub-chemiluminescence detection band 4 of transparent particle, waste liquid suction zones 5 and substrate 6.
Wherein, sample drips district 1 and adopts high porosint, and specifically, sample pad material adopts glass fibre element paper and carries out pretreatment, and pretreatment fluid is large molecule water wetted material.
In enzyme mark monoclonal antibody 1 land 2, enzyme mark thing is alkaline phosphatase (ALP).Alkaline phosphatase has carried out marked by streptavidin.
Enzyme mark monoclonal antibody 1 land 2 adopts glass fibre element made of paper standby, prepares and after carrying out pre-service, drying, then sprays enzyme mark-monoclonal antibody 1, low temperature drying.
Antigen capture band 3 adopts NC film preparation, and be the fixed area of monoclonal antibody 2, monoclonal antibody 2 adopts spraying method to be fixed on NC film.
Optical detection band 4 adopts NC film preparation, and fix by carving rectangle fracture on NC film transparent particle modified, transparent particle selects PVA material to prepare, the homogeneous transparent spheroid of diameter between 100nm ~ 50 μm, its surface adopts carboxyl or hydroxyl hydrophile function group to modify, and is combined with antiantibody and polychlorostyrene benzyl ethylene benzyl Dimethyl Ammonium (BDMQ) stabilizing agent and reinforcing agent.
Waste liquid suction zones 5 adopts strong absorptive material to prepare, and substrate 6 adopts NC film preparation.
As a preferred embodiment of the invention, the invention also discloses the preparation method of above-mentioned chemiluminescence immunoassay examination bar, comprise the following steps:
Step 1, on transparent NC film substrate 6, pre-bonding one deck water wettability thin paper, is stained with NC film (film cuts out rectangle clear area in advance, for the sub-fixed band of reserved transparent particle), immobilized monoclonal antibody 2 on NC film, dry; Glue thieving paper material.
Be fixed in above-mentioned rectangle clear area.
Step 3, on substrate 6 the glass fibre element paper of immobilized enzyme mark monoclonal antibody land 2, spraying monoclonal antibody linked with peroxidase 1, dry.
The glass fibre element paper of step 4, last fixed sample pad 1.
Step 5, slitting, hermetically drying is preserved.
In addition, the invention also discloses another preferred embodiment of preparative chemistry electrochemiluminescent immunoassay examination bar.The all-glass paper being used for sample pad and monoclonal antibody reacting pad is used PBS (phosphoric acid) the damping fluid pre-service drying for standby that contain 0.1%TritonX-100 by pretreatment work.Even application Abl-ALP (alkali phosphatase enzyme mark monoclonal antibody 1) compound on monoclonal antibody reacting pad; Discharge rate is 2 ~ 5 μ l/cm, drying at room temperature.NC film is cut out in advance rectangle (1mm × 3mm) chemical detection band, distance chemiluminescence detection band 5mm place spraying antigen capture band, monoclonal antibody 2 is sprayed on this region, and discharge rate is 1 ~ 2 μ l/cm, drying for standby.PVA-PS composite transparent particulate is selected to modify antiantibody 1 and CDP-Star (two chloro-5-(4-methoxyls { 1,2-dioxetane-3,2 '-(5 '-chlorine) three ring [three thia three ring [3.3.1.13,7] }-4-phenyl phosphate ester) luminous substrate, add the stabilizing agents such as reinforcing agent and bovine serum albumin(BSA) BSA such as Cetylpyridinium chloride trimethyl ammonium CTAC.Then on transparent plastic substrate PET base plate, be stained with NC film and thieving paper, then paste upper monoclonal antibody reacting pad and sample pad successively, then add the sub-material of the transparent particle modified in chemiluminescence detection band micro-syringe, slitting, sealing is preserved.
Through experimental verification, find that the detection sensitivity of chemiluminescence immunoassay of the present invention examination bar is far above colloid gold test paper of the prior art, can be competent at the quick detection of the extremely low concentration materials such as protein macromolecule in biological sample completely.
The present invention has used preferred embodiment to be described, and preferred embodiment is for illustrative purposes, instead of limitation of the present invention.The present invention can be many modifications and changes on the basis of the above description.Therefore, within the scope of the appended claims, the present invention can have is not other above-mentioned implementation.Such as: the preparation method etc. of the combination of particulate kind (as square frame-shaped etc.), chemical illuminating reagent, chemiluminescence band.
Above-described specific embodiment; object of the present invention, technical scheme and beneficial effect are further described; be understood that; the foregoing is only specific embodiments of the invention; be not limited to the present invention; within the spirit and principles in the present invention all, any amendment made, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (10)
1. a chemiluminescence immunoassay examination bar, comprising:
Sample drips district, adopts hydrophilic material manufacture, and testing sample drips in this region;
Trapping region, is fixed with antibody, antigen and/or the antiantibody that can catch molecules detected in testing sample in described trapping region;
Chemiluminescence detection district, transparent particle is fixed with in described detection zone, described transparent particle sub-surface is modified with can molecules detected is combined in described testing sample antibody, antigen and/or antiantibody, and chemical luminous substrate, chemiluminescence stabilizing agent and/or reinforcing agent, due to transparent particle in conjunction with amplification, realize the Sensitive Detection of the faint light of testing sample;
Wherein, described transparent particle is prepared by transparent organic polymer material.
2. chemiluminescence immunoassay examination bar as claimed in claim 1, it is characterized in that, the homogeneous transparent spheroid that described transparent particle is diameter between 100nm ~ 50 μm, described transparent particle be selected from polyvinyl alcohol microparticles, polyvinyl pyrrolidone microballoon, chitosan microball, PVA-PS complex microsphere, polyvinyl alcohol-gelatine microballoon, polypropylene microballoon, polyethyleneglycol diacrylate microballoon, PEG-DA/HEMA gel copolymer microballoon one of at least; And
Described transparent particle sub-surface has been carried out carboxyl, hydroxyl or sulfydryl hydrophile function group and has been modified.
3. chemiluminescence immunoassay examination bar as claimed in claim 1, is characterized in that, described chemical luminous substrate is luminol and derivant, different luminol and derivant thereof, 1,2-dichloroethane luminophor or dioxetane.
4. chemiluminescence immunoassay examination bar as claimed in claim 1, it is characterized in that, when described chemiluminescence immunoassay examination strip adoption HRP luminophor, described chemiluminescence stabilizing agent and reinforcing agent select fluorescein, thiazole, iodophenol, 6-hydroxybenzothiazole, 4-iodobenzene boric acid and/or bromjophenol blue; When described chemiluminescence immunoassay examination strip adoption ALP luminescence system, described chemiluminescence stabilizing agent and reinforcing agent select polychlorostyrene benzyl ethylene benzyl Dimethyl Ammonium BDMQ, TMQ and/or BSA.
5. chemiluminescence immunoassay examination bar as claimed in claim 1, it is characterized in that, when described transparent particle sub-surface is combined with antiantibody, described antiantibody has carried out biotin labeling.
6. chemiluminescence immunoassay examination bar as claimed in claim 1, it is characterized in that, described trapping region comprises enzyme mark monoclonal antibody 1 land and monoclonal antibody 2 trapping region, described enzyme mark monoclonal antibody 1 land is fixed with enzyme mark-monoclonal antibody 1, antigen in testing sample is combined with described enzyme mark-monoclonal antibody 1, forms " enzyme mark-antibody 1-antigen " compound; Described monoclonal antibody 2 trapping region is fixed with monoclonal antibody 2, and the antigen in testing sample is combined with described monoclonal antibody 2, forms " enzyme mark-antibody 1-Ag-Ab 2 " compound; And
The unnecessary enzyme mark-monoclonal antibody 1 in described enzyme mark monoclonal antibody 1 land to be reacted and luminous along with the luminous substrate of transparent particle sub-surface in the flowing of liquid and described chemiluminescence detection district, thus in described testing sample, the concentration of antigen and the luminous intensity in described chemiluminescence detection district are negative correlation.
7. chemiluminescence immunoassay examination bar as claimed in claim 6, it is characterized in that, the enzyme mark thing in described enzyme mark monoclonal antibody 1 land is horseradish peroxidase HR or alkaline phosphatase.
8. a preparation method for chemiluminescence immunoassay examination bar, comprises the following steps:
Step 1, on a transparent substrate, pre-bonding one deck water wettability thin paper, is stained with glass fibre element film, immobilized monoclonal antibody 2 on described glass fibre element film, dry;
Step 2, on nitrocellulose filter, the fixing transparent particle modified is sub; Described transparent particle is prepared by transparent organic polymer material, be selected from polyvinyl alcohol microparticles, polyvinyl pyrrolidone microballoon, chitosan microball, PVA-PS complex microsphere, polyvinyl alcohol-gelatine microballoon, polypropylene microballoon, polyethyleneglycol diacrylate microballoon, PEG-DA/HEMA gel copolymer microballoon one of at least, diameter is between 100nm ~ 50 μm, and described transparent particle sub-surface has been carried out carboxyl, hydroxyl or sulfydryl hydrophile function group and modified;
The glass fibre element paper of step 3, immobilized enzyme mark monoclonal antibody land, spraying monoclonal antibody linked with peroxidase 1, dry;
Step 4, fixed sample pad glass fibre element paper.
Step 5, slitting, hermetically drying is preserved.
9. the preparation method of chemiluminescence immunoassay examination bar as claimed in claim 8, it is characterized in that, fix described transparent particle by carving rectangle white space on described nitrocellulose filter in step 2, or fix described transparent particle by described transparent particle being painted on described nitrocellulose filter surface.
10. the preparation method of chemiluminescence immunoassay examination bar as claimed in claim 8, it is characterized in that, the enzyme mark thing in the mark of enzyme described in step 3 monoclonal antibody 1 land is alkaline phosphatase, and described alkaline phosphatase has carried out marked by streptavidin.
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| CN106918693A (en) * | 2015-12-24 | 2017-07-04 | 上海微柯力高分子材料有限公司 | A kind of microballoon micro-fluidic detection technology based on specific dimensions microballoon and plastic basis material |
| CN108414736A (en) * | 2018-02-08 | 2018-08-17 | 江苏三联生物工程有限公司 | A method of the antibody test object modified using chemiluminescent enhancement |
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