CN111024948A - Immunochromatographic test strip and preparation method thereof - Google Patents
Immunochromatographic test strip and preparation method thereof Download PDFInfo
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- CN111024948A CN111024948A CN201911378115.3A CN201911378115A CN111024948A CN 111024948 A CN111024948 A CN 111024948A CN 201911378115 A CN201911378115 A CN 201911378115A CN 111024948 A CN111024948 A CN 111024948A
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- 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
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
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- 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
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- 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
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57473—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving carcinoembryonic antigen, i.e. CEA
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- 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
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
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- 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
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/585—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with a particulate label, e.g. coloured latex
Abstract
The invention belongs to the technical field of biological detection, and discloses an immunochromatographic test strip which comprises a sample pad, a combination pad and an analysis membrane provided with a detection line and a quality control line, wherein a first mixed antibody marked by a marker is sprayed on the combination pad, a second monoclonal antibody for identifying one site of an object to be detected is coated on the detection line, and a second mixed antibody is coated on the quality control line, wherein the first mixed antibody comprises a third antibody and a first monoclonal antibody for identifying the other site of the object to be detected, and the second mixed antibody comprises an antibody resisting the third antibody and an antibody resisting the first monoclonal antibody. The invention also discloses a preparation method of the immunochromatographic test strip. The invention provides an immunochromatographic test strip and a preparation method thereof, which can optimize the linear range and the precision of the immunochromatographic test strip.
Description
Technical Field
The invention relates to the technical field of biological detection, in particular to an immunochromatographic test strip and a preparation method thereof.
Background
The traditional double-antibody sandwich immunochromatographic test strip comprises a water absorption pad, a nitrocellulose membrane (NC membrane), a quality control line (C line), a detection line (T line), a combination pad, a sample pad and a PVC bottom plate. The sample is added from the sample addition port, flows through the sample pad, the combination pad and the NC membrane, and flows to one side of the absorbent paper. When the sample passes through the bonding pad, the sample to be detected and the immune probe (fluorescent substance, colloidal gold and quantum dot) marked with the antibody on the bonding pad are subjected to immunoreaction to form a first compound. And (3) continuously surging the compound I and the unreacted labeling substance forward under the capillary force of chromatography, and capturing the compound I by a detection line (T line) antibody coated and fixed on the NC membrane to form a compound II. The unreacted labeled substance is partially captured by the antibody coated and fixed on the quality control line (C line) to form a third complex. The first compound which does not react with the T line and the rest of the labeled substance which does not react with the C line are absorbed by the absorbent paper at the tail end. And detecting the signal sizes on the T line and the C line by using a marker analyzer, wherein the concentration of the sample to be detected is in direct proportion to the signal ratio T/C.
The current common technology has two schemes, one quality control line is an anti-mouse system, also called a dependent quality control system, and only one labeled antibody on the corresponding combination pad can identify the mouse IgG monoclonal antibody of the antigen site of the substance to be detected. The T line is coated with another mouse IgG monoclonal antibody capable of identifying other sites of the antigen of the substance to be detected, and the C line is coated with goat anti-mouse polyclonal antibody. The unique label on the pad is captured at the T-line and C-line, respectively, to form a label signal. Along with the increase of the concentration of the sample to be detected object, the T line signal in the signal ratio is increased, and the C line signal is reduced, so that the T/C ratio increasing amplitude is enlarged, and the linear range is enlarged. However, when a high concentration sample is detected in this mode, the C-line signal is usually reduced to a lower level, so that when there is a small difference in C-signal between different strips, a large fluctuation in T/C is caused, and it is difficult to obtain a good detection precision.
The other quality control line is a rabbit antibody or chicken antibody system, also called an independent quality control system, and the corresponding labeled antibodies on the binding pad are two, namely a mouse IgG monoclonal antibody capable of identifying the antigen site of the object to be detected and rabbit IgG or chicken IgY capable of being bound with the C-line antibody. The T line is coated with another mouse IgG monoclonal antibody capable of identifying other sites of the antigen of the substance to be detected, and the C line is coated with goat anti-rabbit or goat anti-chicken or rabbit anti-chicken. The T value signal increases along with the concentration of the substance to be detected from low to high, and the C value signal is basically unchanged. The advantage of this mode is that high value precision is better, but the linear range is narrower because the C signal is unchanged only by the increase of the T signal.
Based on the above situation, there is a need to design an immunochromatographic test strip capable of solving the above problems.
Disclosure of Invention
The invention aims to: the immunochromatographic test strip can optimize the linear range and the precision of the immunochromatographic test strip.
Another object of the invention is: the preparation method of the immunochromatographic test strip can optimize the linear range and the precision of the immunochromatographic test strip.
To achieve the purpose, on one hand, the invention adopts the following technical scheme:
an immunochromatographic test strip comprises a sample pad, a combination pad and an analysis membrane provided with a detection line and a quality control line, wherein the combination pad is sprayed with a first mixed antibody marked by a marker, the detection line is coated with a second monoclonal antibody for identifying a site of an object to be detected, the quality control line is coated with a second mixed antibody, and the first mixed antibody and the second mixed antibody are combined to form the immunochromatographic test strip,
the first mixed antibody comprises a third antibody and a first monoclonal antibody for recognizing another site of the sample, and the second mixed antibody comprises an antibody against the third antibody and an antibody against the first monoclonal antibody.
Further, the third antibody is a rabbit IgG antibody, and the anti-third antibody is an anti-rabbit IgG antibody; or, the third antibody is an anti-rabbit IgG antibody, and the anti-third antibody is a rabbit IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, and the anti-third antibody is an anti-chicken IgY antibody; alternatively, the first and second electrodes may be,
the third antibody is an anti-chicken IgY antibody, and the anti-third antibody is a chicken IgY antibody.
Further, the first monoclonal antibody is a mouse IgG monoclonal antibody, and the antibody against the first monoclonal antibody is an anti-mouse antibody.
Further, the second monoclonal antibody is a murine IgG monoclonal antibody.
Further, the third antibody is a rabbit IgG antibody, the anti-third antibody is a goat anti-rabbit IgG antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a rabbit IgG antibody, the anti-third antibody is a goat anti-rabbit IgG antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-rabbit IgG antibody, the anti-third antibody is a rabbit IgG antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-rabbit IgG antibody, the anti-third antibody is a rabbit IgG antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a goat anti-chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a goat anti-chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a rabbit anti-chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a rabbit anti-chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a rabbit anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a rabbit anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody.
Furthermore, the immunochromatographic test strip further comprises a water absorption pad and a bottom plate, wherein the sample pad, the combination pad, the analysis membrane and the water absorption pad are sequentially overlapped and arranged on the bottom plate.
Further, the sample pad is made of a glass cellulose membrane, a polyester cellulose membrane or filter paper; and/or the presence of a gas in the gas,
the material of the combination pad is a glass cellulose membrane, a polyester cellulose membrane or filter paper; and/or the presence of a gas in the gas,
the analysis membrane is made of a nitrocellulose membrane; and/or the presence of a gas in the gas,
the bottom plate is made of a PVC plate; and/or the presence of a gas in the gas,
the marker is one or more of fluorescent microspheres, magnetic microspheres, colloidal gold, colored latex microspheres, quantum dots, fluorescein, biotin-avidin or a microsphere amplification system.
On the other hand, the invention adopts the following technical scheme:
a preparation method of an immunochromatographic test strip comprises the following steps:
sequentially overlapping and arranging a sample pad, a combination pad, an analysis membrane and a water absorption pad on a bottom plate, wherein the analysis membrane is provided with a detection line and a quality control line, the combination pad is sprayed with a first mixed antibody marked by a marker, the detection line is coated with a second monoclonal antibody for identifying a site of an object to be detected, and the quality control line is coated with a second mixed antibody,
the first mixed antibody comprises a third antibody and a first monoclonal antibody for recognizing another site of the sample, and the second mixed antibody comprises an antibody against the third antibody and an antibody against the first monoclonal antibody.
Further, the third antibody is a rabbit IgG antibody, and the anti-third antibody is an anti-rabbit IgG antibody; or, the third antibody is an anti-rabbit IgG antibody, and the anti-third antibody is a rabbit IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, and the anti-third antibody is an anti-chicken IgY antibody; alternatively, the first and second electrodes may be,
the third antibody is an anti-chicken IgY antibody, and the anti-third antibody is a chicken IgY antibody.
Further, the first monoclonal antibody is a mouse IgG monoclonal antibody, the antibody against the first monoclonal antibody is an anti-mouse antibody, and the second monoclonal antibody is a mouse IgG monoclonal antibody.
Further, the third antibody is a rabbit IgG antibody, the anti-third antibody is a goat anti-rabbit IgG antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a rabbit IgG antibody, the anti-third antibody is a goat anti-rabbit IgG antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-rabbit IgG antibody, the anti-third antibody is a rabbit IgG antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-rabbit IgG antibody, the anti-third antibody is a rabbit IgG antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a goat anti-chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a goat anti-chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a rabbit anti-chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a rabbit anti-chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a rabbit anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a rabbit anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody.
Further, the step of coating the first mixed antibody labeled with the labeling substance on the bonding pad is as follows:
s11: soaking the base material of the bonding pad in a treating fluid, and drying;
s12: preparing a first mixed antibody-label complex;
s13: and (5) spraying the compound prepared in the step (S12) on the bonding pad base material obtained in the step (S11), and drying to obtain the bonding pad.
Further, the sample pad is made of a glass cellulose membrane, a polyester cellulose membrane or filter paper; and/or the presence of a gas in the gas,
the material of the combination pad is a glass cellulose membrane, a polyester cellulose membrane or filter paper; and/or the presence of a gas in the gas,
the analysis membrane is made of a nitrocellulose membrane; and/or the presence of a gas in the gas,
the bottom plate is made of a PVC plate; and/or the presence of a gas in the gas,
the formula of the treating fluid is 0.01-0.05M tris-HCL, 0.5-10% BSA, 1.0-15% trehalose, 1.0-15% sucrose, 0.1-3% PVP, 0.05-5% TW-20, 0.01-0.5% Proclin300, and the pH is 6.0-8.5; and/or the presence of a gas in the gas,
the marker is one or more of fluorescent microspheres, magnetic microspheres, colloidal gold, colored latex microspheres, quantum dots, fluorescein, biotin-avidin or a microsphere amplification system.
Further, the step of coating a second mixed antibody on the quality control line is as follows:
and diluting the second mixed antibody by using a buffer solution, scratching the second mixed antibody on an analysis membrane by using a film scratching instrument, and drying to obtain a quality control line.
The invention has the beneficial effects that: the immunochromatographic test strip and the preparation method thereof are provided, and the linear range and the precision of the immunochromatographic test strip can be optimized. When the immunochromatographic test strip is used for detection, a first monoclonal antibody-to-be-detected object-second monoclonal antibody form a compound, which is embodied as a T-line signal; the first monoclonal antibody forms a first complex with an antibody against the first monoclonal antibody, which is a signal of the C-line; the third antibody forms a second complex with the anti-third antibody, which is reflected in another signal of the C-line. The two C-line signals are superimposed, with the first complex signal decreasing with increasing detection concentration and the second complex signal remaining constant with increasing detection concentration. When a sample with low concentration to high concentration is detected, the C line signal of the invention has not only the reduced amplitude of the C line signal of the dependent quality control system but also the C line signal of the independent quality control system is not changed, thus having better detection precision than the dependent quality control system and better detection linear range than the independent quality control system, and simultaneously, because the multi-reactance of the quality control line is cheaper, the cost is not increased too much, and the scheme is easy to realize.
Drawings
Various other advantages and benefits will become apparent to those of ordinary skill in the art upon reading the following detailed description of the preferred embodiments. The drawings are only for purposes of illustrating the preferred embodiments and are not to be construed as limiting the invention. Also, like reference numerals are used to refer to like parts throughout the drawings. In the drawings:
FIG. 1 is a schematic structural diagram of an immunochromatographic test strip of the present invention;
FIG. 2 is a graph showing T/C values according to concentration for examples and comparative examples of the present invention.
Wherein, the detection device comprises 1-a sample pad, 2-a combination pad, 3-a detection line, 4-a quality control line, 5-an analysis membrane, 6-a water absorption pad and 7-a bottom plate.
Detailed Description
The present invention will be described in further detail with reference to specific examples. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers. The various chemicals used in the examples are commercially available.
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 in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
The immunochromatographic test strip of the invention is shown in figure 1, and comprises a sample pad 1, a combination pad 2 and an analysis membrane 5 provided with a detection line 3 and a quality control line 4, wherein the combination pad 2 is sprayed with a first mixed antibody marked by a marker, the detection line 3 is coated with a second monoclonal antibody for identifying a site of an object to be detected, the quality control line 4 is coated with a second mixed antibody, wherein,
the first mixed antibody comprises a third antibody and a first monoclonal antibody for recognizing another site of the test object, and the second mixed antibody comprises an antibody against the third antibody and an antibody against the first monoclonal antibody.
Specifically, the third antibody and the first monoclonal antibody in the first mixed antibody are mixed with each other, and the antibody against the third antibody and the antibody against the first monoclonal antibody in the second mixed antibody are mixed with each other, and the mixing ratio is not particularly limited, so that the intensity requirement of the detection signal can be satisfied. Except that the first monoclonal antibody for identifying the site of the object to be detected is monoclonal antibody, the other antibodies can be monoclonal antibody or polyclonal antibody. The first monoclonal antibody and the third antibody are each labeled with a labeling substance.
When the immunochromatographic test strip is used for detection, a first monoclonal antibody-to-be-detected object-second monoclonal antibody form a compound, which is embodied as a T-line signal; the first monoclonal antibody forms a first complex with an antibody against the first monoclonal antibody, which is a signal of the C-line; the third antibody forms a second complex with the anti-third antibody, which is reflected in another signal of the C-line. The two C-line signals are superimposed, with the first complex signal decreasing with increasing detection concentration and the second complex signal remaining constant with increasing detection concentration. The invention has the advantages that the C line signal of the invention has not only the reduced amplitude of the C line signal of the dependent quality control system but also the C line signal of the independent quality control system is not changed, so the invention has better detection precision than the dependent quality control system and better detection linear range than the independent quality control system, and simultaneously, the cost is not increased too much because the multi-reactance of the quality control line is cheaper, and the scheme is easy to realize.
In some preferred embodiments, the third antibody is a rabbit IgG antibody, and the anti-third antibody is an anti-rabbit IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is an anti-rabbit IgG antibody, and the anti-third antibody is a rabbit IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, and the antibody for resisting the third antibody is an anti-chicken IgY antibody; alternatively, the first and second electrodes may be,
the third antibody is an anti-chicken IgY antibody, and the antibody for resisting the third antibody is the chicken IgY antibody.
In some preferred embodiments, the first monoclonal antibody is a murine IgG mab and the antibody directed against the first monoclonal antibody is an anti-murine antibody.
In some preferred embodiments, the second monoclonal antibody is a murine IgG mab.
In some preferred embodiments, the third antibody is a rabbit IgG antibody, the anti-third antibody is a goat anti-rabbit IgG antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a rabbit IgG antibody, the anti-third antibody is a goat anti-rabbit IgG antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-rabbit IgG antibody, the anti-third antibody is a rabbit IgG antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-rabbit IgG antibody, the anti-third antibody is a rabbit IgG antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a goat anti-chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a goat anti-chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a rabbit anti-chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a rabbit anti-chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a rabbit anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a rabbit anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody.
As shown in figure 1, the immunochromatographic test strip further comprises a water absorption pad 6 and a bottom plate 7, wherein the sample pad 1, the combination pad 2, the analysis membrane 5 and the water absorption pad 6 are sequentially overlapped and arranged on the bottom plate 7.
The sample pad is made of a glass cellulose membrane, a polyester cellulose membrane or filter paper; the material of the combining pad is glass cellulose membrane, polyester cellulose membrane or filter paper; the material of the analysis membrane is a nitrocellulose membrane; the bottom plate is made of a PVC plate. In some preferred embodiments, the sample pad and the conjugate pad are both made of glass cellulose membranes.
The marker is one or more of fluorescent microspheres, magnetic microspheres, colloidal gold, colored latex microspheres, quantum dots, fluorescein, biotin-avidin or a microsphere amplification system. In some preferred embodiments, the label is a conventional fluorescent microsphere.
The invention also provides a preparation method of the immunochromatographic test strip, which comprises the following steps:
the sample pad 1, the combination pad 2, the analysis membrane 5 and the absorbent pad 6 are sequentially overlapped on a bottom plate 7, the analysis membrane 5 is provided with a detection line 3 and a quality control line 4, the combination pad 2 is sprayed with a first mixed antibody marked by a marker, the detection line 3 is coated with a second monoclonal antibody for identifying a site of an object to be detected, the quality control line 4 is coated with a second mixed antibody, wherein,
the first mixed antibody comprises a third antibody and a first monoclonal antibody for recognizing another site of the sample, and the second mixed antibody is an antibody against the third antibody and an antibody against the first monoclonal antibody.
In some preferred embodiments, the first mixed antibody, the second mixed antibody, the first monoclonal antibody, the second monoclonal antibody, the third antibody, the anti-third antibody, and the anti-first monoclonal antibody are the same as those in the immunochromatographic test strip, and are not described herein again.
The step of coating the conjugate pad with the first mixed antibody labeled with the labeling substance is as follows:
s11: soaking the base material of the bonding pad in a treating fluid, and drying;
s12: preparing a first mixed antibody-label complex;
s13: and (4) spraying the compound prepared in the step (S12) on the bonding pad base material obtained in the step (S11), and drying to obtain the bonding pad fixed with the marker.
In some embodiments, step S12 includes:
cleaning microspheres: washing the fluorescent microspheres with MES solution with pH value of 5.5-7.0, centrifuging at 8000-15000 r/min for 20min, removing supernatant, and repeating the operation for 1-3 times;
and (3) activation: re-suspending the fluorescent microspheres by MES solution, adding an activating agent EDC, and activating for 10-30 min;
labeling the antibody: resuspending the activated fluorescent microspheres by buffer solutions of PB, boric acid or HEPPS and the like with the pH value of 6.0-9.0, adding a mouse IgG antibody and a rabbit or chicken system antibody of a substance to be detected, carrying out shake reaction for 1-3h at room temperature, then centrifuging at the speed of 8000-15000 r/min for 10-30min, and removing supernatant;
sealing the microspheres: resuspending the marked microspheres with a marking buffer solution, adding 1% BSA, shaking and sealing for 0.5-2h, centrifuging at 8000-15000 r/min for 10-30min, and removing supernatant;
and (3) preserving the microspheres: resuspending the microspheres with a microsphere washing buffer (20mM Tris-HCl, 0.9% NaCl (wt/vol), 0.1% Tween-20(vol/vol) and 0.05% Proclin-300(vol/vol), pH8.0), centrifuging at 8000-15000 r/min for 10-30min, discarding the supernatant, repeating the step for 2 times, and finally adding a microsphere preservation solution (10-100mM Tris-HCl, 0.1-1% BSA (wt/vol), 0.5-10% trehalose (wt/vol), 0.5-10% sucrose (wt/vol), 0.1-1% PVP (wt/vol), 0.05-0.5% Tween-20(vol/vol) and 0.05% Proclin-300(vol/vol), carrying out ultrasonic dispersion at pH 8.5), and carrying out resuspension at 4 ℃ in a sealed manner.
The sample pad is made of a glass cellulose membrane, a polyester cellulose membrane or filter paper, the sample pad is soaked in the treatment solution and dried for later use, and the treatment process of the sample pad is a conventional technical means in the field and is not repeated herein;
the material of the combining pad is glass cellulose membrane, polyester cellulose membrane or filter paper;
the material of the analysis membrane is a nitrocellulose membrane;
the bottom plate is made of a PVC plate;
the formula of the treating fluid is 0.01-0.05M tris-HCL, 0.5-10% BSA, 1.0-15% trehalose, 1.0-15% sucrose, 0.1-3% PVP, 0.05-5% TW-20, 0.01-0.5% Proclin300, the pH is 6.0-8.5, the preparation process of the treating fluid is a conventional technical means in the field, and the description is omitted;
the marker is one or more of fluorescent microspheres, magnetic microspheres, colloidal gold, colored latex microspheres, quantum dots, fluorescein, biotin-avidin or a microsphere amplification system.
The steps of coating the second mixed antibody on the quality control line are as follows:
and diluting the second mixed antibody by using a buffer solution, scratching the second mixed antibody on an analysis membrane by using a film scratching instrument, and drying to obtain a quality control line. The buffer solution can be any buffer solution that is conventional in the art and will not be described herein. The detection line is prepared by adopting conventional technical means, and is not described in detail herein.
The first embodiment is as follows: carcinoembryonic antigen CEA fluorescence immunochromatographic test strip (Mixed C line quality control system)
The PVC test paper is characterized in that a sample pad, a combination pad, an NC film and a water absorption pad are sequentially overlapped on a PVC base plate, the width of each functional area is 1-2mm, then the PVC base plate and attached materials are cut into test paper strips with the width of 4mm, a card shell is filled, and a sample adding area and a window test area are arranged on the card shell.
Wherein, the sample pad is glass fiber membrane CB08, contains mouse IgG antibody and chicken IgY antibody of europium ion fluorescence microballon mark carcinoembryonic antigen CEA on the combination pad, and PVC bottom plate material is polyvinyl chloride PVC board, and the analysis membrane is the NC membrane of material Sartorius CN140 area backing, contains on the NC membrane: the T line (detection line) is coated with another CEA monoclonal antibody capable of being specifically combined with the CEA antigen to be detected, the C line (quality control line) is coated with a mixed polyclonal antibody of goat anti-mouse IgG and rabbit anti-chicken IgY, and the europium ion fluorescent microspheres contain carboxyl with the particle size of 200 nm.
The test strip of the embodiment can be prepared by the following method:
(1) coating T line and C line antibodies on NC membrane
① fixing and sticking the NC film on the middle part of the PVC bottom plate;
② diluting CEA mouse IgG monoclonal antibody 1 (second monoclonal antibody) to 1mg/ml with coating buffer solution (20mM PB, pH7.4) as the coating solution of detection line (T line), and coating solution of quality control line (C line) containing goat anti-mouse IgG (anti-first monoclonal antibody) and rabbit anti-chicken IgY (anti-third antibody) mixed polyclonal antibody (second mixed antibody) diluted with coating buffer solution (20mM PB, pH7.4) at concentrations of 0.5mg/ml and 0.5mg/ml, respectively, and then scratching the T and C line solution onto NC film with gold-spraying film scratching instrument to fix the antibody;
③ drying the ② NC film in an oven at 37 deg.C for 4h, adding desiccant, and storing under sealed condition.
(2) Sample pad preparation
The sample pad material is glass fiber membrane CB08, width 13mm, sample pad buffer: 20mM Tris-HCl, 0.5% Tween-20, 2% BSA, 5% sucrose, 1.5% PVP, pH 8.5.
② the sample pad is soaked in the buffer solution for 1 hour, then taken out, and dried in an oven at 37 ℃ for 4 hours.
(3) Bond pad preparation
① cutting bonding pad, bonding pad material glass fiber 6614, cutting the bonding pad into 8 mm/300 mm width;
② A marker diluent and a conjugate pad treatment solution were prepared, 2 formulations were the same, 0.015Mtris-HCl, BSA 0.5% (wt/vol), trehalose 2.0% (wt/vol), sucrose 5% (wt/vol), PVP 0.25% (wt/vol), TW-200.05% (vol/vol), Proclin 3000.01% (vol/vol), pH 8.5.
③ the conjugate pad was soaked in the treatment solution for 1 hour, then taken out and dried in an oven at 37 ℃ for 4 hours.
(4) Mouse IgG monoclonal antibody and chicken IgY antibody mark of CEA
Cleaning microspheres: taking 50 mu L of europium ion fluorescent microspheres (solid content is 1%) in a 2mL centrifugal tube, performing 300W ultrasonic dispersion treatment for 1min by using 1mL MES buffer solution (50mM, pH5.5), centrifuging for 15min at 15000rpm, removing supernatant, and repeating the step for 3 times;
and (3) activation: adding 0.5mL MES buffer (50mM, pH5.5) for ultrasonic resuspension, sequentially adding 40 μ g and 20 μ g of Sulfo-NHS and EDC, vortex mixing, incubating at 37 ℃ for 15min with a shaker, centrifuging at 15000rpm for 15min, discarding the supernatant, and repeating the step for 2 times;
labeling the antibody: adding 0.5mL PB labeling buffer (20mM, pH7.4) and performing 300W ultrasonic dispersion treatment for 1min, adding 40 μ g CEA mouse IgG monoclonal antibody 2 (first monoclonal antibody), and performing shaking table reaction at 37 ℃ for 2 h;
sealing the microspheres: adding 40uL BSA (10% by mass, diluted with pure water) microsphere blocking buffer, reacting for 1h at 37 ℃ by a shaking table, centrifuging for 15min at 15000rpm, and removing the supernatant;
and (3) preserving the microspheres: 0.5mL of microsphere washing buffer (20mM Tris-HCl, 0.9% NaCl (wt/vol), 0.1% Tween-20(vol/vol) and 0.05% Proclin-300(vol/vol), pH8.0) was added for ultrasonic resuspension, centrifuged at 15000rpm for 15min, the supernatant was discarded, the procedure was repeated 2 times, and finally 0.25mL of microsphere preservation solution (25mM Tris-HCl, 1% BSA (wt/vol), 5% trehalose (wt/vol), 5% sucrose (wt/vol), 1% PVP (wt/vol), 0.05% Tween-20(vol/vol) and 0.05% Proclin-300(vol/vol), pH8.5 was added for ultrasonic resuspension, and the mixture was sealed and preserved at 4 ℃.
The labeling process of the chicken IgY antibody (third antibody) is consistent with that of the mouse IgG monoclonal antibody of CEA.
(5) Conjugate pad spray microsphere coupled antibody Complex (first Mixed antibody)
Diluting the CEA-fluorescent microsphere conjugate and the chicken IgY-fluorescent microsphere conjugate to 0.5mg/mL and 0.1mg/mL by using a microsphere diluent, spraying the mixed labeling microsphere solution on the bonding pad treated by the bonding pad treatment solution by using a gold spraying and membrane scratching instrument, and drying for 7 hours by using an air-blast drying oven.
(6) CEA test strip assembly
A sample pad, a combination pad sprayed with microsphere coupling antibody compounds, an NC film coated with antibodies and a water absorption pad are sequentially overlapped on a polyvinyl chloride (PVC) bottom plate, the width of each functional area is 1-2mm, then the PVC bottom plate and attached materials are cut into test strips with the width of 4mm, and the test strips are loaded into a card shell. And drying agent is added into the detection card placed in the aluminum foil bag for sealing and preservation.
(7) Detection of
The concentrations of the quality control substances of the detection objects are respectively set as 5.8ng/ml, 92.1ng/ml, 418.5ng/ml and 582.4ng/ml, each concentration quality control substance is tested for 10 times, and the change conditions of precision CV and T/C value of each concentration along with the increase of the concentration are calculated. The results are shown in Table 1.
Table 1: example one test result
Comparative example one: carcinoembryonic antigen CEA fluorescence immunochromatographic test strip (dependent quality control system)
This comparative example is the same as the first majority of the procedure, except that:
(1) NC Membrane preparation, coating solution of the control line (C line) contained only goat anti-mouse IgG polyclonal antibody diluted with coating buffer (20mM PB, pH 7.4).
(4) Only labeled CEA mouse IgG monoclonal antibody, not labeled chicken IgY antibody.
(5) Only the CEA-fluorescent microsphere conjugate is sprayed on the bonding pad, and the chicken IgY-fluorescent microsphere conjugate is not sprayed on the bonding pad.
Similarly, the concentrations of the quality control substances to be detected are respectively set to be 5.8ng/ml, 92.1ng/ml, 418.5ng/ml, 582.4ng/ml and 500ng/ml, each concentration quality control substance is tested for 10 times, and the change of precision CV and T/C value of each concentration along with the increase of the concentration is calculated. The results are shown in table 2 and fig. 2.
Table 2: comparative example 1 test results
Comparative example two: carcinoembryonic antigen CEA fluorescence immunochromatographic test strip (independent quality control system)
This comparative example is the same as the first majority of the procedure, except that:
(1) NC Membrane preparation, coating solution of the quality control line (line C) contained only rabbit anti-chicken IgY polyclonal antibody diluted with coating buffer (20mM PB, pH 7.4).
Similarly, the concentrations of the quality control substances to be detected are respectively set to be 5.8ng/ml, 92.1ng/ml, 418.5ng/ml, 582.4ng/ml and 500ng/ml, each concentration quality control substance is tested for 10 times, and the change of precision CV and T/C value of each concentration along with the increase of the concentration is calculated. The results are shown in table 3 and fig. 2.
Table 3: comparative example II test results
In the aspect of detecting the linear range, as can be seen from fig. 2, the T/C signal increases with the increase of the sample concentration, the medium-high concentration, the non-independent quality control system has the best signal amplification, the mixed C-line quality control system has the next time, the signal amplification of the independent quality control system is very small, and the high-value concentration cannot be distinguished.
In terms of precision, as can be seen from tables 1, 2 and 3, the precision CV of the mixed C-line quality control system and the independent quality control system is greater than 10% instead of the high concentration point of the independent quality control system, because the C-line signal of the dependent quality control system is more reduced when detecting a high concentration sample, on one hand, the accuracy of capturing a high signal by the detecting instrument is reduced when capturing a low signal, and on the other hand, the signal value is determined by T/C, and obviously, when the C signal fluctuates at a low level, a large fluctuation of the T/C value is easily caused.
Therefore, the mixed C-line quality control system can control good precision under the condition of ensuring that the high-concentration sample is detected with a certain degree of distinction, and obtains beneficial detection effect compared with an independent quality control system and a non-independent quality control system. Since the above-mentioned other antibody combinations of the first mixed antibody and the second mixed antibody have the same principle as the antibody combination in this embodiment, it should be understood by those skilled in the art that the same experimental conclusion as this embodiment can be obtained when the immunochromatographic test strip made of other antibody combinations is used for detection, and thus the detailed description is omitted here.
The mixed C-line quality control system in the invention refers to a detection system of the immunochromatographic test strip, and the name is only used for convenience of expression and is distinguished from a traditional independent quality control system and a non-independent quality control system.
The parts not referred to in the present invention are the same as or can be implemented using the prior art.
Finally, it should be noted that: in the description of the present invention, the technical terms "first" and "second" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance. In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art. The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims (11)
1. An immunochromatographic test strip comprising a sample pad, a conjugate pad, and an analytical membrane provided with a detection line and a quality control line, wherein the conjugate pad is sprayed with a first mixed antibody labeled with a labeling substance, the detection line is coated with a second monoclonal antibody identifying a site of an object to be detected, and the quality control line is coated with a second mixed antibody, wherein,
the first mixed antibody comprises a third antibody and a first monoclonal antibody for recognizing another site of the sample, and the second mixed antibody comprises an antibody against the third antibody and an antibody against the first monoclonal antibody.
2. The immunochromatographic test strip according to claim 1,
the third antibody is a rabbit IgG antibody, and the anti-third antibody is an anti-rabbit IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is an anti-rabbit IgG antibody, and the anti-third antibody is a rabbit IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, and the anti-third antibody is an anti-chicken IgY antibody; alternatively, the first and second electrodes may be,
the third antibody is an anti-chicken IgY antibody, and the anti-third antibody is a chicken IgY antibody.
3. The immunochromatographic test strip of claim 1 or 2, wherein the first monoclonal antibody is a murine IgG monoclonal antibody, and the antibody against the first monoclonal antibody is an anti-murine antibody.
4. The immunochromatographic test strip of claim 3, wherein the second monoclonal antibody is a murine IgG monoclonal antibody.
5. The immunochromatographic test strip according to claim 4,
the third antibody is a rabbit IgG antibody, the anti-third antibody is a goat anti-rabbit IgG antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a rabbit IgG antibody, the anti-third antibody is a goat anti-rabbit IgG antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-rabbit IgG antibody, the anti-third antibody is a rabbit IgG antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-rabbit IgG antibody, the anti-third antibody is a rabbit IgG antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a goat anti-chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a goat anti-chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a rabbit anti-chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a chicken IgY antibody, the anti-third antibody is a rabbit anti-chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a goat anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a rabbit anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a goat anti-mouse IgG antibody; alternatively, the first and second electrodes may be,
the third antibody is a rabbit anti-chicken IgY antibody, the anti-third antibody is a chicken IgY antibody, and the anti-mouse antibody is a rabbit anti-mouse IgG antibody.
6. The immunochromatographic test strip according to claim 1, further comprising a water absorption pad and a bottom plate, wherein the sample pad, the binding pad, the analysis membrane and the water absorption pad are sequentially overlapped and disposed on the bottom plate.
7. The immunochromatographic test strip according to claim 6, wherein the sample pad is made of a glass cellulose membrane, a polyester cellulose membrane or filter paper; and/or the presence of a gas in the gas,
the material of the combination pad is a glass cellulose membrane, a polyester cellulose membrane or filter paper; and/or the presence of a gas in the gas,
the analysis membrane is made of a nitrocellulose membrane; and/or the presence of a gas in the gas,
the bottom plate is made of a PVC plate; and/or the presence of a gas in the gas,
the marker is one or more of fluorescent microspheres, magnetic microspheres, colloidal gold, colored latex microspheres, quantum dots, fluorescein, biotin-avidin or a microsphere amplification system.
8. The method for preparing the immunochromatographic test strip of any one of claims 1 to 7, comprising the steps of:
sequentially overlapping and arranging a sample pad, a combination pad, an analysis membrane and a water absorption pad on a bottom plate, wherein the analysis membrane is provided with a detection line and a quality control line, the combination pad is sprayed with a first mixed antibody marked by a marker, the detection line is coated with a second monoclonal antibody for identifying a site of an object to be detected, and the quality control line is coated with a second mixed antibody,
the first mixed antibody comprises a third antibody and a first monoclonal antibody for recognizing another site of the sample, and the second mixed antibody comprises an antibody against the third antibody and an antibody against the first monoclonal antibody.
9. The method for preparing the immunochromatographic test strip according to claim 8, wherein the step of spraying the first mixed antibody labeled with a labeling substance on the conjugate pad is as follows:
s11: soaking the base material of the bonding pad in a treating fluid, and drying;
s12: preparing a first mixed antibody-label complex;
s13: and (5) spraying the compound prepared in the step (S12) on the bonding pad base material obtained in the step (S11), and drying to obtain the bonding pad.
10. The method for preparing an immunochromatographic test strip according to claim 9,
the sample pad is made of a glass cellulose membrane, a polyester cellulose membrane or filter paper; and/or the presence of a gas in the gas,
the material of the combination pad is a glass cellulose membrane, a polyester cellulose membrane or filter paper; and/or the presence of a gas in the gas,
the analysis membrane is made of a nitrocellulose membrane; and/or the presence of a gas in the gas,
the bottom plate is made of a PVC plate; and/or the presence of a gas in the gas,
the formula of the treating fluid is 0.01-0.05M tris-HCL, 0.5-10% BSA, 1.0-15% trehalose, 1.0-15% sucrose, 0.1-3% PVP, 0.05-5% TW-20, 0.01-0.5% Proclin300, and the pH is 6.0-8.5; and/or the presence of a gas in the gas,
the marker is one or more of fluorescent microspheres, magnetic microspheres, colloidal gold, colored latex microspheres, quantum dots, fluorescein, biotin-avidin or a microsphere amplification system.
11. The method for preparing the immunochromatographic test strip according to claim 8, wherein the step of coating the second mixed antibody on the quality control line is as follows:
and diluting the second mixed antibody by using a buffer solution, scratching the second mixed antibody on an analysis membrane by using a film scratching instrument, and drying to obtain a quality control line.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911378115.3A CN111024948A (en) | 2019-12-27 | 2019-12-27 | Immunochromatographic test strip and preparation method thereof |
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| CN111323580A (en) * | 2020-04-24 | 2020-06-23 | 东莞市东阳光诊断产品有限公司 | Immunochromatographic test strip and preparation method thereof |
| CN111912992A (en) * | 2020-08-28 | 2020-11-10 | 河北国高生物科技有限公司 | Preparation method of triple-joint detection and determination kit for IV type collagen, laminin and III type procollagen amino-terminal peptide |
| CN112630423A (en) * | 2020-12-31 | 2021-04-09 | 杭州福德敏生物技术有限公司 | Wheat immunofluorescence detection test strip and application |
| CN112698028A (en) * | 2020-12-22 | 2021-04-23 | 海卫特(广州)医疗科技有限公司 | Immunochromatography pad and preparation method and application thereof |
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| CN111912992A (en) * | 2020-08-28 | 2020-11-10 | 河北国高生物科技有限公司 | Preparation method of triple-joint detection and determination kit for IV type collagen, laminin and III type procollagen amino-terminal peptide |
| CN112698028A (en) * | 2020-12-22 | 2021-04-23 | 海卫特(广州)医疗科技有限公司 | Immunochromatography pad and preparation method and application thereof |
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| CN116539872A (en) * | 2023-04-25 | 2023-08-04 | 上海润普生物技术有限公司 | Fluorescent immunochromatography detection kit for quantitative platelet antibody and preparation method thereof |
| CN116539872B (en) * | 2023-04-25 | 2024-02-09 | 上海润普生物技术有限公司 | Fluorescent immunochromatography detection kit for quantitative platelet antibody and preparation method thereof |
| CN116990510A (en) * | 2023-09-26 | 2023-11-03 | 聚诚(北京)生物科技有限责任公司 | Adenovirus antigen reagent strip for detecting ocular secretion based on colloidal gold method and preparation method thereof |
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