CN117269483A - Detection test paper based on agglutination reaction and test method thereof - Google Patents
Detection test paper based on agglutination reaction and test method thereof Download PDFInfo
- Publication number
- CN117269483A CN117269483A CN202311558869.3A CN202311558869A CN117269483A CN 117269483 A CN117269483 A CN 117269483A CN 202311558869 A CN202311558869 A CN 202311558869A CN 117269483 A CN117269483 A CN 117269483A
- Authority
- CN
- China
- Prior art keywords
- detection
- chromatographic
- signal
- antibody
- test strip
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000012360 testing method Methods 0.000 title claims abstract description 94
- 238000001514 detection method Methods 0.000 title claims abstract description 92
- 230000004520 agglutination Effects 0.000 title claims abstract description 56
- 238000006243 chemical reaction Methods 0.000 title claims abstract description 50
- 238000010998 test method Methods 0.000 title claims abstract description 20
- 239000000463 material Substances 0.000 claims abstract description 53
- 239000000427 antigen Substances 0.000 claims abstract description 31
- 239000007788 liquid Substances 0.000 claims abstract description 27
- 102000036639 antigens Human genes 0.000 claims abstract description 26
- 108091007433 antigens Proteins 0.000 claims abstract description 26
- 239000003550 marker Substances 0.000 claims abstract description 21
- 239000011259 mixed solution Substances 0.000 claims abstract description 8
- 238000011210 chromatographic step Methods 0.000 claims abstract description 5
- 239000000243 solution Substances 0.000 claims abstract description 4
- 239000003154 D dimer Substances 0.000 claims description 28
- 108010052295 fibrin fragment D Proteins 0.000 claims description 24
- 238000000034 method Methods 0.000 claims description 24
- 239000011148 porous material Substances 0.000 claims description 18
- 239000004005 microsphere Substances 0.000 claims description 17
- 241000287828 Gallus gallus Species 0.000 claims description 14
- 239000004793 Polystyrene Substances 0.000 claims description 13
- 229920002223 polystyrene Polymers 0.000 claims description 13
- 241000283973 Oryctolagus cuniculus Species 0.000 claims description 11
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 10
- 238000009792 diffusion process Methods 0.000 claims description 9
- 238000004587 chromatography analysis Methods 0.000 claims description 8
- 108010090804 Streptavidin Proteins 0.000 claims description 7
- 210000003743 erythrocyte Anatomy 0.000 claims description 5
- 230000036046 immunoreaction Effects 0.000 claims description 3
- 101800000407 Brain natriuretic peptide 32 Proteins 0.000 claims description 2
- 102400000667 Brain natriuretic peptide 32 Human genes 0.000 claims description 2
- 101800002247 Brain natriuretic peptide 45 Proteins 0.000 claims description 2
- 108010074051 C-Reactive Protein Proteins 0.000 claims description 2
- 102100032752 C-reactive protein Human genes 0.000 claims description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 2
- 102000004889 Interleukin-6 Human genes 0.000 claims description 2
- 108090001005 Interleukin-6 Proteins 0.000 claims description 2
- 241001494479 Pecora Species 0.000 claims description 2
- 108010048233 Procalcitonin Proteins 0.000 claims description 2
- 102000054727 Serum Amyloid A Human genes 0.000 claims description 2
- 108700028909 Serum Amyloid A Proteins 0.000 claims description 2
- 102000004903 Troponin Human genes 0.000 claims description 2
- 108090001027 Troponin Proteins 0.000 claims description 2
- 229910052799 carbon Inorganic materials 0.000 claims description 2
- 230000007423 decrease Effects 0.000 claims description 2
- 229940100601 interleukin-6 Drugs 0.000 claims description 2
- HPNRHPKXQZSDFX-OAQDCNSJSA-N nesiritide Chemical compound C([C@H]1C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CSSC[C@@H](C(=O)N1)NC(=O)CNC(=O)[C@H](CO)NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](CCSC)NC(=O)[C@H](CCCCN)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CO)C(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1N=CNC=1)C(O)=O)=O)[C@@H](C)CC)C1=CC=CC=C1 HPNRHPKXQZSDFX-OAQDCNSJSA-N 0.000 claims description 2
- CWCXERYKLSEGEZ-KDKHKZEGSA-N procalcitonin Chemical compound C([C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)NCC(O)=O)[C@@H](C)O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCSC)NC(=O)[C@H]1NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(N)=O)NC(=O)CNC(=O)[C@@H](N)CSSC1)[C@@H](C)O)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 CWCXERYKLSEGEZ-KDKHKZEGSA-N 0.000 claims description 2
- 230000035945 sensitivity Effects 0.000 abstract description 8
- 238000005259 measurement Methods 0.000 abstract 2
- 238000012113 quantitative test Methods 0.000 abstract 1
- 238000001035 drying Methods 0.000 description 21
- 239000012528 membrane Substances 0.000 description 14
- 230000008569 process Effects 0.000 description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 238000010521 absorption reaction Methods 0.000 description 8
- 230000000903 blocking effect Effects 0.000 description 7
- 230000009471 action Effects 0.000 description 6
- 239000007853 buffer solution Substances 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 238000002791 soaking Methods 0.000 description 6
- 238000011161 development Methods 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 230000003321 amplification Effects 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 239000011248 coating agent Substances 0.000 description 4
- 238000000576 coating method Methods 0.000 description 4
- 238000003317 immunochromatography Methods 0.000 description 4
- 229940127121 immunoconjugate Drugs 0.000 description 4
- 238000003199 nucleic acid amplification method Methods 0.000 description 4
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 3
- 241000283707 Capra Species 0.000 description 3
- 229920001030 Polyethylene Glycol 4000 Polymers 0.000 description 3
- 229920001213 Polysorbate 20 Polymers 0.000 description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 3
- 229930006000 Sucrose Natural products 0.000 description 3
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 3
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 3
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 3
- 238000012827 research and development Methods 0.000 description 3
- 238000005507 spraying Methods 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- 239000004094 surface-active agent Substances 0.000 description 3
- 238000004364 calculation method Methods 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 238000004445 quantitative analysis Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 230000003313 weakening effect Effects 0.000 description 2
- 230000004523 agglutinating effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 239000011162 core material Substances 0.000 description 1
- 230000001808 coupling effect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000003365 glass fiber Substances 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 238000012764 semi-quantitative analysis Methods 0.000 description 1
- 230000000405 serological effect Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54386—Analytical elements
- G01N33/54387—Immunochromatographic test strips
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/82—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a precipitate or turbidity
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
- G01N33/537—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with separation of immune complex from unbound antigen or antibody
- G01N33/539—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with separation of immune complex from unbound antigen or antibody involving precipitating reagent, e.g. ammonium sulfate
- G01N33/541—Double or second antibody, i.e. precipitating antibody
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/82—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a precipitate or turbidity
- G01N2021/825—Agglutination
Landscapes
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plasma & Fusion (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The application specifically discloses a detection test paper based on an agglutination reaction and a test method thereof. A test method of detection test paper based on signal attenuation comprises a pretreatment step, a chromatography step and a detection step; introducing a primary antibody marked by a signal marker into the sample mixed solution, thereby obtaining a chromatographic solution; introducing a bispecific antibody into the chromatographic liquid before the chromatographic liquid reaches the signal detection area, and capturing an agglutinated material in the chromatographic liquid by the bispecific antibody, wherein the agglutinated material is a material capable of undergoing an agglutination reaction; and reading an actual measurement signal value belonging to the signal marker, comparing the actual measurement signal value with a standard signal value of the signal marker, and estimating the content of the target antigen in the detection sample based on a comparison result. The application has the advantages of improving the sensitivity of the immunochromatographic test paper and improving the accuracy of qualitative, semi-quantitative and quantitative tests.
Description
Technical Field
The invention relates to the technical field of in-vitro diagnosis, in particular to a test paper based on an agglutination reaction and a test method thereof.
Background
The immunochromatography technology is mainly completed by means of an immunochromatography test strip, is simple and convenient to operate, rapid in detection, strong in specificity, good in stability and convenient to carry, and has been widely applied to the fields of clinical diagnosis, food safety, drug detection, environmental pollution and the like.
At present, the immunochromatographic test paper on the market comprises colloidal gold immunochromatographic test paper, fluorescent immunochromatographic test paper and the like, most of the test strips on the market are quantitative detection test strips matched with a quantitative analysis instrument, and the test strips are all detected through the increase of test line signals; based on the principle of signal amplification, the traditional immunochromatography test paper has a large gray area when performing qualitative or semi-quantitative detection, and is easy to cause false negative or false positive results when being clinically used, so that the accuracy of the test is reduced.
Disclosure of Invention
In order to reduce the test ash area of the immunochromatographic test paper and improve the sensitivity and the qualitative and quantitative accuracy of the immunochromatographic test paper, the application provides a detection test paper based on an agglutination reaction and a test method thereof.
In a first aspect, the present application provides a test method for a test strip based on an agglutination reaction, which adopts the following technical scheme:
a test method of detection test paper based on agglutination reaction comprises the following steps:
pretreatment: preparing a sample mixed solution containing a detection sample, and introducing a primary antibody marked by a signal marker into the sample mixed solution, thereby obtaining a chromatographic solution;
chromatography step: allowing the chromatographic liquid to reach a signal detection area through chromatography, wherein the signal detection area is provided with a secondary antibody for capturing the primary antibody, and a bispecific antibody is introduced into the chromatographic liquid before the chromatographic liquid reaches the signal detection area, wherein the bispecific antibody can specifically capture a target antigen, and the bispecific antibody can specifically capture an agglutinated material in the chromatographic liquid, and the agglutinated material is a material capable of undergoing agglutination reaction;
the detection step comprises: detecting the signal detection area, reading an actually measured signal value belonging to the signal marker, comparing the actually measured signal value with a standard signal value of the signal marker, and estimating the content condition of the target antigen in the detection sample based on a comparison result, wherein the standard signal value is a signal value measured by the signal marker under the standard concentration.
Before the chromatographic liquid reaches the color development area, adding an agglutination antibody, capturing a target antigen by the agglutination antibody to generate agglutination reaction, wherein a gelled material obtained by the agglutination reaction can move towards the position close to the color development area along with the progress of the chromatographic process, the particle size of the gelled material obtained by the agglutination reaction is gradually increased along with the progress of the chromatographic process, the gelled material with larger particle size gradually blocks available pore channels for the chromatographic liquid and the gelled material to pass through in the process of moving towards the color development area, and the concentration of the primary antibody and the signal marker which are chromatographed to the position of the signal detection area is reduced along with the progress of the chromatographic process, so that the primary antibody and the signal marker which can be combined by the secondary antibody in the signal detection area are reduced, and the intensity of fluorescent signals is reduced; the content of the target antigen in the reaction sample is obtained by comparing with the standard signal value, so that the target antigen is used for preparing a qualitative, quantitative or semi-quantitative detection reagent.
Agglutination is a serological reaction; the particulate antigen binds to the corresponding antibody and, in the presence of the electrolyte, aggregates visible to the naked eye appear over a period of time. Specifically, the agglutination reaction can be classified into a direct agglutination reaction and an indirect agglutination reaction.
Compared with the enrichment of the beacon at the test line in the traditional chromatography method so as to display the optical signal, the detection method of the application has no signal amplification effect as the traditional kit; the test method is characterized in that a pore channel leading to a signal detection area is blocked by gelled materials generated by the agglutination reaction, so that a signal marker cannot smoothly reach the signal detection area, and the process of signal existence is performed; and in the process of blocking the pore channels, a great amount of signal markers can be trapped when one pore channel is blocked, and the blocking degree and the trapped signal markers are not 1:1, therefore, the detection method has a trapping signal amplification mechanism, has narrower detection gray areas and higher sensitivity compared with the immunochromatography method used in the prior art, and has an order of magnitude improvement compared with the sensitivity of the traditional kit after the detection method is applied to the kit.
In addition, the test method does not need to use monoclonal antibodies, and the cost of the reintroduced bispecific antibodies is low, so that the material cost of the test method can be reduced; in addition, the test method does not need to carry out the operation of coupling monoclonal antibodies, so that the production procedures of a kit applying the method can be reduced, and the research and development period is shortened; in the research and development process of the traditional kit, very high manpower and material resources are often required to be input to screening of several core materials such as monoclonal antibodies, so that the final cost of the kit applying the detection method can be reduced by 60% -80% or more compared with that of the traditional method.
Preferably, the case where the target antigen undergoes the agglutination reaction includes:
the target antigen forms a gelled material by direct agglutination reaction with the agglutinated material; and/or, the agglutinated material undergoes indirect agglutination reaction to form a gelled material.
Preferably, the antigen of interest comprises any one of D-dimer, troponin, brain natriuretic peptide, C-reactive protein, procalcitonin, interleukin 6, serum amyloid a, and the like.
Preferably, the agglutinating material comprises any one of red blood cells and polystyrene microspheres; when the agglutination material is polystyrene microsphere, the polystyrene microsphere is coupled with streptavidin or antibody, and the bispecific antibody captures the polystyrene microsphere through immunoreaction with the streptavidin or antibody.
When the agglutination material is red blood cells, the agglutination reaction of the target antigen with the agglutination material is a direct agglutination reaction. When the agglutination material is polystyrene microsphere, the bispecific antibody can be used for capturing polystyrene microsphere by immunoreaction with streptavidin or antibody.
The test method is suitable for detecting all target antigens which can participate in agglutination reaction, and can intercept the signal markers in the chromatographic process under the action of the gelled materials generated by the agglutination reaction only by selecting proper agglutination materials, so that the detection intensity of the signal markers in the signal detection area is reduced, and qualitative and quantitative detection of the target antigens is realized.
In a second aspect, the present application provides a test strip based on an agglutination reaction, which adopts the following technical scheme:
the detection test paper based on the agglutination reaction comprises a sample adding area and a chromatographic area, wherein the sample adding area is connected with the chromatographic area, and a diffusion area and a signal detection area are sequentially arranged on the chromatographic area along the direction far away from the sample adding area;
the sample adding region is provided with a primary antibody marked by a signal marker, the signal detection region is provided with a secondary antibody for capturing the primary antibody, and when the detection test paper is used for detection, chromatographic liquid containing a detection sample enters the diffusion region from the sample adding region and is chromatographed to the signal detection region through the diffusion region;
the detection test paper also comprises a bispecific antibody, wherein the bispecific antibody can specifically capture an agglutinated material in the chromatographic liquid, and the agglutinated material is a material capable of undergoing an agglutination reaction; the bispecific antibody is disposed in the loading zone and/or the diffusion zone.
The agglutination material capable of undergoing agglutination reaction is selected to react with the target antigen to generate a gelled material, so that the concentration of the target antigen transferred to the signal detection area is reduced, the detection of the target antigen is realized by reducing the concentration of the target antigen in the signal detection area without screening specific monoclonal antibodies like detection test paper in the prior art, and the application procedures and the application cost of the detection test paper are reduced; in addition, in the actual operation process, the difficulty of coupling the monoclonal antibody is maximum, the fault tolerance is low, the coupling effect is also the least stable, the stability and the detection accuracy of the detection test paper are not facilitated, and the detection test paper in the application can overcome the problem of the detection instability and obviously improve the detection accuracy.
Preferably, the concentration of the bispecific antibody is 0.1-5 μg/mL
By controlling the concentration of the bispecific antibody, the qualitative analysis of the target antigen can be realized, and simultaneously, the quantitative and semi-quantitative analysis can be realized.
Preferably, the primary antibody comprises any one of chicken IgY and rabbit IgG.
Preferably, the concentration of the primary antibody is 0.1-5 μg/mL.
The concentration of the primary antibody is controlled, so that the primary antibody transferred to the signal detection area can be sufficiently detected by the secondary antibody, and the test sensitivity is improved.
Preferably, the signal marker comprises any one of immunochromatographic markers such as colloidal gold, colloidal carbon, fluorescent microspheres, color microspheres and the like.
The beacon substance used in the test method is easy to obtain in source, and the application cost of the test method is reduced.
Preferably, the secondary antibody comprises any one of rabbit anti-chicken IgY and sheep anti-rabbit IgG.
Preferably, when the primary antibody is chicken IgY, the secondary antibody is rabbit anti-chicken IgY; when the primary antibody is rabbit IgG, the secondary antibody is goat anti-rabbit IgG.
Preferably, the porosity of the chromatographic zone decreases gradually in the chromatographic direction of the chromatographic liquid.
When the porosity of the chromatographic zone is gradually reduced along the chromatographic direction of the chromatographic liquid, the gelled material is gradually blocked in the chromatographic zone along with the progress of the chromatographic process, so that the content of the primary antibody at the signal detection zone is reduced, the light signal weakening amplitude at the test line of the signal detection zone is further enhanced due to the occurrence of the gelled material, and the test effect is remarkable.
Preferably, the pore size of the chromatographic zone is 5-12 μm.
Preferably, the test paper based on the agglutination reaction provided by the application comprises a PVC bottom plate 1, and a sample pad 2, a bonding pad 3, an NC film 4 and a water absorption pad 5 which are arranged on the PVC bottom plate 1, wherein the sample pad 2, the bonding pad 3, the NC film 4 and the water absorption pad 5 are sequentially overlapped in pairs, and the NC film 4 is provided with a test wire 41; the sample pad 2 and the conjugate pad 3 correspond to the sample application region, the NC membrane 4 between the conjugate pad 3 and the test line 41 corresponds to the diffusion region, and the test line 41 corresponds to the signal detection region.
Drawings
Fig. 1 is a schematic structural diagram of a test strip according to various embodiments of the present application.
FIG. 2 is a graph showing the quantitative detection standard of D-dimer in example 1.
Reference numerals illustrate:
1. a PVC bottom plate; 2. a sample pad; 3. a bonding pad; 4. NC film; 41. a test line; 5. a water absorbing pad.
Detailed Description
For a better understanding and implementation, the technical solutions of the present invention will be clearly and completely described below in connection with examples.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
Unless otherwise indicated, all numbers expressing quantities of ingredients, reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term "about". Accordingly, unless indicated to the contrary, the numerical parameters set forth herein are approximations that may vary depending upon the desired properties to be obtained.
As used herein, "and/or" means one or all of the elements mentioned.
The use of "including" and "comprising" herein encompasses both the situation in which only the elements are mentioned and the situation in which other elements not mentioned are present in addition to the elements mentioned.
All percentages in the present invention are by weight unless otherwise indicated.
As used in this specification, the terms "a," "an," "the," and "the" are intended to include "at least one" or "one or more," unless otherwise specified. For example, "a component" refers to one or more components, and thus more than one component may be considered and possibly employed or used in the practice of the embodiments.
Examples
In this example, the substrates of the sample pad 2 and the bonding pad 3 are both glass fiber films, and the NC film 4 is CN140 of dolis germany.
Example 1
The detection test paper based on signal attenuation comprises a PVC bottom plate 1, a sample pad 2, a combination pad 3, an NC film 4 and a water absorption pad 5 which are arranged on the PVC bottom plate 1, wherein the sample pad 2, the combination pad 3, the NC film 4 and the water absorption pad 5 are sequentially overlapped in pairs, and a test wire 41 is arranged on the NC film 4;
soaking the sample pad 2 in PBS buffer solution, adding sucrose PEG4000 and the like, and drying at a constant temperature of 37 ℃ in a drying oven for 4 hours for later use;
adding trehalose, a surfactant S9, tween 20 and the like into the binding pad 3 by using BB buffer solution, soaking, drying, coating bispecific antibody of 0.1-5 mug/mL and chicken IgY-colloidal gold conjugate of 0.1-5 mug/mL after the binding pad is dried, and drying for 4 hours at a constant temperature of 37 ℃ in a drying box for later use;
test line 41 is coated with 0.1-5 μg/mL rabbit anti-chicken IgY and dried in a drying oven at 37 ℃ for 4h for standby
In the test paper, the size of the pore canal on the bonding pad 3 is larger than that of the pore canal in the NC membrane 4, and the diameter of the pore canal on the NC membrane 4 is 5-12 mu m.
A test method of detection test paper based on signal attenuation comprises the following steps:
pretreatment: drop-adding whole blood sample as sample mixture to sample pad 2, wherein D-dimer concentration in sample mixture is 0.00, 0.03, 0.11, 0.52, 2.56, 5.69, 13.42, 19.35, 28.21, 40.87 μg/L, each concentration is tested three times;
chromatography step: the whole blood sample containing the D-dimer is chromatographed from the sample pad 2 to the binding pad 3, and red blood cells in the whole blood sample and the D-dimer are subjected to agglutination reaction under the action of the bispecific antibody so as to form a gelled material with a dense space three-dimensional structure; as the chromatographic reaction proceeds, the gelled material gradually plugs the NC membrane 4 between the conjugate pad 3 and the NC membrane 4 and between the conjugate pad 3 and the test line 41, so that the concentration of the chicken IgY-colloidal gold conjugate that can diffuse to the test line 41 by the chromatographic action is reduced;
the detection step comprises: the chicken IgY-colloidal gold conjugate diffused to the test line 41 can be captured by rabbit anti-chicken IgY, the measured signal value belonging to the chicken IgY-colloidal gold conjugate is read, the measured signal value is compared with the standard signal value of the signal marker, and the content condition of the D-dimer is estimated based on the comparison result.
Specifically, a detection light signal value with the D-dimer concentration of 0.00 mug/L in a sample mixed solution is calibrated as a standard value, the concentration is taken as an abscissa, a scatter diagram is established by taking the average value of three test chromogenic signals as an ordinate, a standard curve is fitted, and the detection limit is as low as 0.026 mug/mL and the linear range is 0.026-28.21 mug/mL through calculation; and comparing the color development optical signal value of the sample mixed liquid containing the D-dimer with the standard value to obtain the specific content of the D-dimer in the sample mixed liquid. The standard curve for quantitative detection of D-dimer in this example is shown in FIG. 2.
Example 2
The detection test paper based on signal attenuation comprises a PVC bottom plate 1, a sample pad 2, a combination pad 3, an NC film 4 and a water absorption pad 5 which are arranged on the PVC bottom plate 1, wherein the sample pad 2, the combination pad 3, the NC film 4 and the water absorption pad 5 are sequentially overlapped in pairs, and a test wire 41 is arranged on the NC film 4;
soaking the sample pad 2 in PBS buffer solution, adding sucrose PEG4000 and the like, and drying at a constant temperature of 37 ℃ in a drying oven for 4 hours for later use;
adding trehalose, a surfactant S9, tween 20 and the like into the binding pad 3 by using BB buffer solution, soaking, drying, spraying polystyrene microspheres coated with streptavidin, 0.1-5 mug/mL of bispecific antibody and 0.1-5 mug/mL of chicken IgY-colloidal gold conjugate after drying the binding pad, and drying for 4 hours at a constant temperature of 37 ℃ in a drying box for later use;
coating 0.1-5 mug/mL rabbit anti-chicken IgY on the test line 41, and drying for 4 hours at the constant temperature of 37 ℃ in a drying box for later use;
in the test paper, the size of the pore canal on the bonding pad 3 is larger than that of the pore canal in the NC membrane 4, and the diameter of the pore canal on the NC membrane 4 is 5-12 mu m.
A test method of detection test paper based on signal attenuation comprises the following steps:
pretreatment: drop serum sample as sample mixture onto sample pad 2, wherein the concentration of D-dimer in the sample mixture is 0.00, 0.03, 0.11, 0.52, 2.56, 5.69, 13.42, 19.35, 28.21, 40.87 μg/L, each concentration is tested three times;
chromatography step: the serum sample containing D-dimer is chromatographed from the sample pad 2 to the binding pad 3, and the polystyrene microsphere is subject to agglutination reaction under the action of the bispecific antibody, specifically, the bispecific antibody and streptavidin are subject to immune reaction to realize the capture of the polystyrene microsphere, so that the agglutination reaction is carried out to form a gelled material with a dense space three-dimensional structure; as the chromatographic reaction proceeds, the gelled material gradually plugs the NC membrane 4 between the conjugate pad 3 and the NC membrane 4 and between the conjugate pad 3 and the test line 41, so that the concentration of the chicken IgY-colloidal gold conjugate that can diffuse to the test line 41 by the chromatographic action is reduced;
the detection step comprises: the chicken IgY-colloidal gold conjugate diffused to the test line 41 can be captured by rabbit anti-chicken IgY, the measured signal value belonging to the chicken IgY-colloidal gold conjugate is read, the measured signal value is compared with the standard signal value of the signal marker, and the content condition of the D-dimer is estimated based on the comparison result.
Specifically, a detection light signal value with the D-dimer concentration of 0.00 mug/L in a sample mixed solution is calibrated as a standard value, the concentration is taken as an abscissa, a scatter diagram is established by taking the average value of three test chromogenic signals as an ordinate, a standard curve is fitted, and the detection limit is as low as 0.027 mug/mL and the linear range is 0.027-28.11 mug/mL through calculation; and comparing the color development optical signal value of the sample mixed liquid containing the D-dimer with the standard value to obtain the content of the D-dimer in the sample mixed liquid.
Comparative example 1
The method comprises the steps that detection test paper in the prior art is used for testing, the detection test paper comprises a PVC bottom plate 1, a sample pad 2, a combination pad 3, an NC film 4 and a water absorption pad 5 which are arranged on the PVC bottom plate 1, the sample pad 2, the combination pad 3, the NC film 4 and the water absorption pad 5 are sequentially overlapped in pairs, and a T line and a C line are sequentially arranged on the NC film 4 along the chromatography direction of a chromatographic liquid;
soaking the sample pad 2 in PBS buffer solution, adding sucrose PEG4000 and the like, and drying at a constant temperature of 37 ℃ in a drying oven for 4 hours for later use;
soaking the bonding pad 3 in BB buffer solution, adding trehalose, surfactant S9, tween 20 and the like, drying, spraying 0.1-5 mug/mL of colloidal gold-labeled D-dimer monoclonal antibody conjugate on the bonding pad by using a film-dividing metal spraying instrument after drying, and drying in a drying oven at a constant temperature of 37 ℃ for 4 hours for later use;
in NC film 4, coating mouse anti-human D dimer monoclonal antibody conjugate on T line, coating 0.1-5 mug/mL goat anti-mouse IgG on C line, drying at 37 deg.C for 4 h;
in the test paper, the size of the pore canal on the bonding pad 3 is larger than that of the pore canal in the NC membrane 4, and the diameter of the pore canal on the NC membrane 4 is 5-12 mu m.
As can be seen from the test, the sample mixture is dripped onto the sample pad 2, and the D-dimer in the sample mixture is combined with the colloidal gold mouse anti-human D-dimer monoclonal antibody conjugate to form a colloidal gold-labeled antibody-antigen complex when passing through the combination pad 3; the colloidal gold-labeled antibody-antigen complex is continuously diffused onto the NC membrane 4 along with the sample mixed solution, is intercepted by a T line (detection line) coated with the mouse anti-human D-dimer monoclonal antibody conjugate, and captures the colloidal gold-labeled antibody-antigen complex to form an immune complex of the colloidal gold-labeled antibody-antigen-coated antibody; the immune complex of the non-intercepted colloidal gold labeled antibody-antigen-coated antibody continues to go upwards and is combined with the goat anti-mouse IgG coated by the C line (quality control line) to indicate that the reaction is completed.
The signal value at the C line is relatively stable and is not influenced by the sample mixed liquid, the signal value at the T line is in direct proportion to the concentration of the D dimer in the sample mixed liquid, the ratio of the signal value at the T line to the signal value at the C line is calculated, and a standard curve of the concentration of the T/C and the D dimer in the sample is established, so that the concentration value of the D dimer in the unknown sample is calculated by detecting the T/C value of the unknown sample.
The detection limit of the D-dimer in this comparative example was found to be as low as 0.1. Mu.g/mL, and the linear range was found to be 0.5-10. Mu.g/mL.
In combination with example 1, comparative example 1 and fig. 1, it can be seen that the test strip of the present application has lower detection limit and detection sensitivity, and a larger detection gray area, because, when the D-dimer in the sample and the red blood cells in the whole blood sample undergo an agglutination reaction under the action of the bispecific antibody, the gelled material gradually moves toward the test line 41 as the chromatographic reaction proceeds, and because the particle size of the gelled material increases and the pore size of the junction between the conjugate pad 3 and the NC film 4 becomes smaller during the chromatographic process, the gelled material is blocked at the junction between the conjugate pad 3 and the NC film 4 and at the NC film 4 between the conjugate pad 3 and the test line 41; with the progress of the chromatographic reaction, the chicken IgY antibody on the binding pad 3 blocks the chromatographic passage because of the gelled material, and the blocking degree is in direct proportion to the content of the target antigen in the sample; the concentration of the chicken IgY antibody-beacon substance transferred to the test line 41 is reduced, thereby reducing the concentration of the chicken IgY antibody-beacon substance to which the rabbit anti-chicken IgY on the test line 41 can bind, and weakening the signal intensity displayed on the test line 41. In addition, in the process that the NC film 4 is blocked, a large amount of beacon substances can be trapped by blocking one hole, and the blocking degree and the trapped amount of the beacon substances are not in a simple 1:1 relation, so that the detection test paper has a trapping signal amplification mechanism, and the detection sensitivity of the scheme is improved by orders of magnitude compared with that of the traditional kit; and the production cost is reduced, a specific monoclonal antibody is not needed to be selected, and the research and development period is shortened.
By combining examples 1-2 with fig. 1, it can be seen that by adopting the test method in the present application, for different target antigens, a suitable bispecific antibody and an agglutination material are selected, so that the agglutination material generated by the agglutination reaction moves along the direction of the chromatographic fluid along with the progress of the chromatographic process, thereby blocking the pore canal at the lap joint of the NC membrane 4 and the binding pad 3 or/and the pore canal in the NC membrane 4 between the binding pad 3 and the test line 41, blocking the primary antibody and the signal marker, and judging the concentration of the target antigen by the intensity of the fluorescent signal displayed at the test line 41, thereby significantly improving the sensitivity of the test paper. Also, for the D-dimer test, the limit of detection in example 2 was similar to that in example 1, and during the specific procedure, the chromatographic background in example 2 was found to be cleaner than in example 1.
The present embodiment is merely illustrative of the present application and is not intended to be limiting, and those skilled in the art, after having read the present specification, may make modifications to the present embodiment without creative contribution as required, but is protected by patent laws within the scope of the claims of the present application.
Claims (12)
1. A chromatography test method based on an agglutination reaction, characterized in that: the method comprises the following steps:
pretreatment: preparing a sample mixed solution containing a detection sample, and introducing a primary antibody marked by a signal marker into the sample mixed solution, thereby obtaining a chromatographic solution;
chromatography step: allowing the chromatographic liquid to reach a signal detection area through chromatography, wherein the signal detection area is provided with a secondary antibody for capturing the primary antibody, and a bispecific antibody is introduced into the chromatographic liquid before the chromatographic liquid reaches the signal detection area, wherein the bispecific antibody can specifically capture a target antigen, and the bispecific antibody can specifically capture an agglutinated material in the chromatographic liquid, and the agglutinated material is a material capable of undergoing agglutination reaction;
the detection step comprises: detecting the signal detection area, reading an actually measured signal value belonging to the signal marker, comparing the actually measured signal value with a standard signal value of the signal marker, and estimating the content condition of the target antigen in the detection sample based on a comparison result, wherein the standard signal value is a signal value measured by the signal marker under the standard concentration.
2. The method for testing chromatography based on the agglutination reaction according to claim 1, wherein: the conditions in which the agglutination of the antigen of interest occurs include:
the target antigen forms a gelled material by direct agglutination reaction with the agglutinated material;
and/or, the agglutinated material undergoes indirect agglutination reaction to form a gelled material.
3. The method for testing chromatography based on the agglutination reaction according to claim 1, wherein: the target antigen comprises any one of D-dimer, troponin, brain natriuretic peptide, C-reactive protein, procalcitonin, interleukin 6, serum amyloid A and the like.
4. A method of testing by chromatography based on the agglutination reaction according to claim 3, wherein: the agglutination material comprises any one of red blood cells and polystyrene microspheres; when the agglutination material is polystyrene microsphere, the polystyrene microsphere is coupled with streptavidin or antibody, and the bispecific antibody can capture the polystyrene microsphere through immunoreaction with the streptavidin or antibody.
5. A test paper based on agglutination reaction, characterized in that: the detection test paper comprises a sample adding area and a chromatographic area, wherein the sample adding area is connected with the chromatographic area, and a diffusion area and a signal detection area are sequentially arranged on the chromatographic area along the direction away from the sample adding area;
the sample adding region is provided with a primary antibody marked by a signal marker, the signal detection region is provided with a secondary antibody for capturing the primary antibody, and when the detection test paper is used for detection, chromatographic liquid containing a detection sample enters the diffusion region from the sample adding region and is chromatographed to the signal detection region through the diffusion region;
the detection test paper also comprises a bispecific antibody, wherein the bispecific antibody can specifically capture an agglutinated material in the chromatographic liquid, and the agglutinated material is a material capable of undergoing an agglutination reaction; the bispecific antibody is disposed in the loading zone and/or the diffusion zone.
6. The test strip of claim 5, wherein the test strip comprises: the concentration of the bispecific antibody is 0.1-5 mug/mL.
7. The test strip of claim 5, wherein the test strip comprises: the primary antibody comprises any one of chicken IgY and rabbit IgG.
8. The test strip of claim 5, wherein the test strip comprises: the concentration of the primary antibody is 0.1-5 mug/mL.
9. The test strip of claim 5, wherein the test strip comprises: the signal marker comprises any one of immunochromatographic markers such as colloidal gold, colloidal carbon, fluorescent microspheres, color microspheres and the like.
10. The test strip of claim 5, wherein the test strip comprises: the secondary antibody comprises any one of rabbit anti-chicken IgY and sheep anti-rabbit IgG.
11. The test strip of claim 5, wherein the test strip comprises: the porosity of the chromatographic zone gradually decreases along the chromatographic direction of the chromatographic liquid.
12. The test strip of claim 5, wherein the test strip comprises: the pore diameter of the chromatographic zone is 5-12 mu m.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311558869.3A CN117269483B (en) | 2023-11-22 | 2023-11-22 | Detection test paper based on agglutination reaction and test method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311558869.3A CN117269483B (en) | 2023-11-22 | 2023-11-22 | Detection test paper based on agglutination reaction and test method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN117269483A true CN117269483A (en) | 2023-12-22 |
| CN117269483B CN117269483B (en) | 2024-03-29 |
Family
ID=89218142
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311558869.3A Active CN117269483B (en) | 2023-11-22 | 2023-11-22 | Detection test paper based on agglutination reaction and test method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117269483B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4963498A (en) * | 1985-08-05 | 1990-10-16 | Biotrack | Capillary flow device |
| WO2006047831A1 (en) * | 2004-11-03 | 2006-05-11 | Agen Biomedical Limited | Detection device and method |
| US20100267065A1 (en) * | 2009-03-25 | 2010-10-21 | Timothy Robert Geiger | Apparatus and methods for analyzing fluid variables |
| US20170226578A1 (en) * | 2014-07-14 | 2017-08-10 | Oxford University Innovation Limited | Measurement of analytes with membrane channel molecules, and bilayer arrays |
| CN115728495A (en) * | 2022-10-20 | 2023-03-03 | 华中农业大学 | Charge-mediated micro-channel current biosensing method based on directional coupling technology and application thereof |
-
2023
- 2023-11-22 CN CN202311558869.3A patent/CN117269483B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4963498A (en) * | 1985-08-05 | 1990-10-16 | Biotrack | Capillary flow device |
| WO2006047831A1 (en) * | 2004-11-03 | 2006-05-11 | Agen Biomedical Limited | Detection device and method |
| US20100267065A1 (en) * | 2009-03-25 | 2010-10-21 | Timothy Robert Geiger | Apparatus and methods for analyzing fluid variables |
| US20170226578A1 (en) * | 2014-07-14 | 2017-08-10 | Oxford University Innovation Limited | Measurement of analytes with membrane channel molecules, and bilayer arrays |
| CN115728495A (en) * | 2022-10-20 | 2023-03-03 | 华中农业大学 | Charge-mediated micro-channel current biosensing method based on directional coupling technology and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117269483B (en) | 2024-03-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR970007079B1 (en) | Determination of ambient concentration of several analytes | |
| EP0516095A2 (en) | Process and device for specific binding assay | |
| CN111198273B (en) | Immunoassay kit for anti-phospholipase A2 receptor autoantibody, preparation method and use method thereof | |
| CN108593919A (en) | A kind of colloidal gold immune chromatography test and its preparation method and application | |
| CN111896750A (en) | Bovine immunoglobulin (IgG) double-antibody sandwich colloidal gold immunochromatographic test strip, preparation method, kit and detection method | |
| CN105988008A (en) | Measurement device, kit and measurement method | |
| CN111896749A (en) | Bovine lactoferrin double-antibody sandwich colloidal gold immunochromatographic test strip, preparation method, kit and detection method | |
| JPH0575396B2 (en) | ||
| Tu et al. | Ultrasensitive heterogeneous immunoassay using photothermal deflection spectroscopy | |
| van Amerongen et al. | Quantitative computer image analysis of a human chorionic gonadotropin colloidal carbon dipstick assay | |
| CN117269483B (en) | Detection test paper based on agglutination reaction and test method thereof | |
| CN106526166A (en) | Rapid detection of lean meat powder in pork | |
| US5583003A (en) | Agglutination assay | |
| CN117031022A (en) | Kit and method for detecting plasmin by fluorescence immunochromatography | |
| EP4286850A1 (en) | Immunological assay method | |
| EP0643836B1 (en) | Agglutination assays and kits employing colloidal dyes | |
| GB2590230A (en) | Methods and kits for detection of 11-dehydro-thromboxane B2 | |
| CN113113079B (en) | Method for identifying hook effect in quantitative immunochromatography test | |
| CN111896743A (en) | Fluorescence immunochromatography test strip and preparation method and application thereof | |
| WO2011064910A1 (en) | Immunity measurement method | |
| JP2000258418A (en) | Measuring method by using immuno-chromatography and test body analytical tool used therein | |
| CN116413445A (en) | Detection card, kit and detection method for detecting total thyroxine content | |
| JP5714791B2 (en) | Non-specific reaction inhibitor | |
| IE903439A1 (en) | Agglutination assay | |
| JP3328053B2 (en) | Determination of antibody or antigen concentration by immunoagglutination |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |