CN116466072A - Dry-type fluorescence immunochromatography detection card, kit and detection method for quantitatively detecting procalcitonin and heparin binding protein - Google Patents
Dry-type fluorescence immunochromatography detection card, kit and detection method for quantitatively detecting procalcitonin and heparin binding protein Download PDFInfo
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Classifications
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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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
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- 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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/533—Production of labelled immunochemicals with fluorescent label
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- G—PHYSICS
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- 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
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
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- 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
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/575—Hormones
- G01N2333/585—Calcitonins
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
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Abstract
The invention discloses a dry-type fluorescence immunochromatography detection card, a kit and a detection method for quantitatively detecting procalcitonin and heparin binding proteins; the test strip in the detection card comprises a sample pad, a combination pad, a reaction membrane, an absorption pad and a bottom plate; the reaction film is stuck to the middle part of one surface of the bottom plate, and the left side and the right side of the reaction film are respectively laminated, buckled and lapped with one side of the bonding pad and one side of the absorption pad; one side edge of the sample pad is laminated, buckled and lapped on the other side edge of the bonding pad, and the sample pad and the absorption pad are respectively arranged at two ends of the bottom plate; two detection lines and a quality control line which are parallel to each other are arranged in the middle of the reaction film, and the two detection lines are adjacent; the test strip is based on double-antibody sandwich fluorescence immunity lateral chromatography, improves the quantitative detection sensitivity of procalcitonin/heparin binding protein, has simple and convenient operation of the kit, and can obtain accurate results by directly adding samples, and has the advantages of simple operation flow, short detection time, low detection cost and the like.
Description
Technical Field
The invention relates to the field of biomedical detection, in particular to a dry fluorescent immunochromatography detection card and a kit for quantitatively detecting procalcitonin and heparin binding proteins.
Background
Procalcitonin (PCT) is a procalcitonin precursor protein synthesized by thyroid C cells, consisting of 116 amino acids and having a molecular weight of 13kDa. When the organism is stimulated by bacterial endotoxin, PCT can be rapidly increased in a short time, and is currently considered as an important index for identifying bacterial and viral infection in early clinical stage and judging prognosis of severe patients. PCT is generally considered to reflect the activity level of systemic inflammatory response, and its influencing factors include the size and type of the infected organ, the kind of bacteria, the degree of inflammation, the condition of immune response, and the like. Even without bacterial infection or bacterial lesions, PCT levels are elevated when the body is subjected to shock, immune response, organ failure, but weaker than when it is subjected to endotoxin infection. Heparin Binding Protein (HBP) is an important granule protein secreted by polymorphonuclear leucocytes, also an acute phase protein, can be released along with the aggregation of neutrophils after body infection, and has chemotactic property, sterilization activity and inflammation regulation effect. As a novel inflammation marker, the early disease condition can be evaluated, the dynamic development of the disease condition can be judged, and the clinical application guidance has higher clinical value and important significance.
Currently, commonly used detection methods of procalcitonin/heparin binding proteins include radioimmunoassay, chemiluminescent method, transmission immunonephelometry, dry immunofluorescent method, electrochemiluminescent method and colloidal gold colorimetric method, and these methods have advantages and disadvantages, and are specifically as follows:
1) The principle of radioimmunoassay is to use the principle of labeling an antigen or antibody with a radionuclide, binding the antigen or antibody to be tested, and forming an antigen-antibody complex. A competitive RIA, labeled with an antigen, based on the principle that an unknown antigen ag+a labeled antigen ag+a known quantitative antibody Ab labeled antigen-antibody complex agab+an unknown antigen-antibody complex agab+a free labeled antigen Ag (removed) (more unknown antigen, less complex bound by the labeled antigen and the quantitative antibody, and represented as a negative correlation), and another non-competitive RIA, labeled with an antibody, generating a sandwich-like antibody-antigen-antibody complex, removing the free labeled antibody, and measuring the radiometric count of the solid-phase antibody-antigen-labeled antibody complex; this method is time consuming and can result in cross-reactions and false positive reactions.
2) The chemiluminescent label immunoassay is also called chemiluminescent immunoassay, and uses a double monoclonal antibody to be respectively combined with molecules, so that the method can eliminate cross reaction. Chemiluminescent immunoassay devices comprise two components, namely an immunoreaction system and a chemiluminescent assay system. The chemiluminescence analysis system is characterized in that a chemiluminescent substance is catalyzed by a catalyst and oxidized by an oxidant to form an excited intermediate, when the excited intermediate returns to a stable ground state, photons are emitted at the same time, and the light quantum yield is measured by a luminescence signal measuring instrument. The immune reaction system is to directly label a luminescent substance (an excited state intermediate generated under the excitation of a reactant) on an antigen (chemiluminescent immunoassay) or an antibody (immunochemical immunoassay), or to act an enzyme on a luminescent substrate. The core detector in the chemiluminescence immunoassay instrument is a photomultiplier, single photon detection is carried out, the photomultiplier is transmitted to an amplifier, high-voltage current is added for amplification, the amplifier converts analog current into digital current, and a light-emitting signal is transmitted to a computer through an R232 data line and calculated to obtain a clinical result. The chemiluminescent immunoassay technology has the advantages of high sensitivity, wide linear dynamic range, stable result and small error; the disadvantages are that specialized chemiluminescent instruments need to be paired, and most of the devices are closed systems, reagents and instruments cannot be used commonly among different manufacturers, the cost is high, the operation is time-consuming, and operators have to have special operation skills.
3) The transmission immune turbidimetry is that antigen and antibody form antigen-antibody complex in a certain buffer solution, when light rays penetrate through a reaction solution, the reflection and absorption of the antigen-antibody complex particles in the solution to the light rays cause the reduction of transmitted light, and the more the antigen-antibody complex is, the less the transmitted light is, the absorbance can be used for representing, but the accuracy is low.
4) Dry immunofluorescence is a dry chromatography based on immunofluorescence quantitative detection techniques: mixing detection buffer solution and a blood sample in a detection bottle, wherein fluorescent marked anti-antibody in the buffer solution and antigen in blood form an antigen-antibody complex, when the mixed sample is placed on a sample of a reaction plate and is diffused onto a test strip of a nitrocellulose matrix through capillary action, the complex is captured by the antibody on the test strip, the signal intensity of the fluorescent antibody reflects the captured quantity, and the concentration in a blood sample can be reflected through the treatment of an iCHROMA Reader immunofluorescence analyzer. The method has the advantages of simple preparation, low operation cost, poor activity and influence on the detection result.
5) Electrochemiluminescence is a phenomenon in which a voltage or current signal of a certain waveform is applied to an electrode to cause a reaction between products of electrolytic reaction or with coexisting components in a system to generate chemiluminescence. The sensitivity is high, the linear range is wide, the instrument and equipment are simple, the automation is easy to realize, most of instruments are imported, the cost is relatively high, and the laboratory requirement is high.
6) The colloidal gold colorimetric method adopts the technology of colloidal gold to form gold-labeled antigen-antibody complexes, and the method is quick, simple and easy to operate, but cannot accurately judge the content.
Disclosure of Invention
In view of the above problems, one of the problems to be solved by the present invention is to provide a dry-type fluorescence immunochromatography detection card based on fluorescence immunochromatography and added with fluorescent microspheres, which can realize simple operation and rapid quantitative detection of procalcitonin and heparin binding proteins
The invention aims to solve the second problem and provide a dry fluorescent immunochromatography kit for quantitatively detecting procalcitonin and heparin binding proteins.
The first technical scheme of the invention is as follows:
a dry fluorescent immunochromatographic assay card for quantitatively detecting procalcitonin and heparin binding proteins, which is characterized by comprising a test strip and a card shell adapted to accommodate the test strip; wherein:
the test strip comprises a sample pad, a combination pad, a reaction membrane, an absorption pad and a bottom plate; the bottom plate is in a strip structure, the reaction film is stuck to the middle part of one surface of the bottom plate, and the left side and the right side of the reaction film are respectively laminated, buckled and lapped with one side of the bonding pad and one side of the absorption pad; one side edge of the sample pad is laminated, buckled and overlapped on the other side edge of the bonding pad, and the sample pad and the absorption pad are respectively arranged at two ends of the bottom plate; the binding pad is coated with procalcitonin/heparin binding protein marked by fluorescent microspheres; two detection lines and a quality control line which are parallel to each other are arranged in the middle of the reaction film, and the two detection lines are adjacent; the detection line is coated with procalcitonin/heparin binding protein antigen; the quality control line is coated with chicken IgY monoclonal antibody;
the clamping shell comprises a containing cavity, a sample adding hole and an observation window; the sample adding hole and the observation window are respectively communicated with the accommodating cavity; when the test strip is placed in the accommodating cavity of the cartridge, the sample pad is exposed through the sample adding hole, and the reaction membrane is exposed through the observation window.
In one embodiment, in the dry-type fluorescence immunochromatography detection card, the binding pad is coated with a procalcitonin/heparin binding protein antibody which is marked by fluorescent microspheres and recognizes different epitopes and a goat anti-chicken IgY monoclonal antibody marked by the fluorescent microspheres.
In one embodiment, in the dry-type fluorescence immunochromatography detection card, the separation distance between the detection line and the quality control line is 5±0.5mm.
In one embodiment, in the dry-type fluorescence immunochromatography detection card, the coating liquid for coating the procalcitonin/heparin binding protein antigen and the chicken IgY monoclonal antibody is a mixed liquid comprising 0.01M sucrose and 0.01M PBS buffer solution with pH of 7.4, and the concentration of the mixed liquid is 1mg/ml.
In one embodiment, in the dry-type fluorescence immunochromatography detection card, the sample pad is a glass cellulose membrane which is dried after being soaked in a surfactant buffer solution.
In one embodiment, the dry fluorescent immunochromatographic assay card comprises BSA, tween-20 having a pH of 8.0.+ -. 0.05, and boric acid-borax buffer having a pH of 0.2M.
In one embodiment, in the dry-type fluorescence immunochromatography detection card, the reaction membrane is made of nitrocellulose membrane.
In one embodiment, in the dry-type fluorescent immunochromatographic assay card, the bonding pad is made of a glass cellulose membrane.
The second solution of the present invention is to provide a dry fluorescent immunochromatography kit for quantitatively detecting procalcitonin and heparin binding proteins, which comprises any one of the above dry fluorescent immunochromatography detection cards, a sample diluent and a chip; reagent standard curve information is stored in the chip; the sample diluent is used for diluting serum, plasma and/or whole blood samples to be detected.
The invention also provides a method for detecting the content of procalcitonin and heparin binding protein in serum, plasma and/or whole blood by using the kit, which comprises the following steps:
firstly, adding serum, plasma and/or whole blood samples into a sample diluent, and uniformly mixing to obtain a mixed sample;
secondly, adding the mixed sample to a sample pad of the test strip, and displaying a detection result by a detection card within 10-15 minutes;
finally, after the detection card in the kit is inserted into a matched immunofluorescence analyzer, the concentration of procalcitonin and heparin binding protein in serum, plasma and/or whole blood samples is detected.
The immunochromatography detection card and the kit for quantitatively detecting procalcitonin and heparin binding protein (PCT/HBP) provided by the invention are based on a double-antibody sandwich method and fluorescence immunity lateral chromatography, provide a high-sensitivity procalcitonin/heparin binding protein immunochromatography quantitative detection kit, are simple and convenient to operate, and can obtain accurate results by directly adding samples. When the invention is used, the detection sensitivity is improved, and the invention has the advantages of simple operation flow, short detection time, low detection cost and the like.
Drawings
FIG. 1 is a schematic diagram of a test strip provided with a test card according to the present invention;
fig. 2 is a schematic diagram of a card housing structure of a test card according to the present invention.
Detailed Description
The preferred embodiments of the present invention will be described in further detail with reference to the accompanying drawings.
The invention provides an immunochromatography detection card and a kit for quantitatively detecting procalcitonin and heparin binding protein (PCT/HBP); the detection card comprises a test paper strip and a card shell for accommodating the test paper strip; specifically, the following is described.
As shown in FIG. 1, a PCT/HBP immunochromatography quantitative detection test strip 10 comprises a bottom plate 15 made of PVC material and provided with a sample pad 11, a binding pad 12, a reaction membrane 13 and an absorption pad 14 in sequence. One side of the absorption pad 14 and one side of the bonding pad 12 are respectively overlapped and lapped on two opposite sides of the reaction film 13, a detection area is formed on the surface of the reaction film 13, and two detection lines 16 coated with fluorescent markers and a quality control line (C line) 17 coated with chicken IgY monoclonal antibodies are fixed in the detection area; the two detection lines (T lines) 16 and the quality control lines are arranged in parallel at intervals, and the two detection lines 16 are adjacent; the detection line 16 and the quality control line 17 are respectively parallel to the short side edge of the bottom plate 15; wherein, the binding pad 12 is coated with PCT/HBP antibodies which are marked by fluorescent microspheres and recognize different epitopes, sheep anti-chicken IgY monoclonal antibodies marked by the fluorescent microspheres, and procalcitonin/heparin binding protein antigens marked by the fluorescent microspheres.
The sample pad 11 is a glass fiber membrane which is dried after being soaked in a surfactant buffer solution; wherein the active agent buffer solution is boric acid-borax buffer solution containing BSA and Tween-20,0.2M with pH value of 8.0+/-0.05.
The reaction membrane is made of a nitrocellulose membrane, and the bonding pad is made of a glass cellulose membrane; the absorbent pad is made of H-1 absorbent paper.
As shown in fig. 2, the cartridge 20 includes a receiving chamber 21, a loading hole 23, and an observation window 22; the sample adding hole 23 and the observation window 22 are respectively communicated with the accommodating cavity 21; when the test strip 10 is placed in the accommodating cavity 21 of the cartridge 20, the sample pad 12 is exposed through the sample application hole 23, and the reaction membrane 13 is exposed through the observation window 22. Namely, the bottom plate 15, the sample pad 11, the bonding pad 12, the reaction membrane 13 and the absorption pad 14 of the test strip 10 are all disposed in the cartridge 20, and an observation window 22 is disposed on the surface of the cartridge 20 corresponding to the reaction membrane 13, and a sample loading hole 23 is disposed on the surface of the cartridge 20 corresponding to the sample pad 11.
The invention also provides a dry-type fluorescence immunochromatography kit for quantitatively detecting procalcitonin and heparin binding proteins, which comprises the dry-type fluorescence immunochromatography detection card, a sample diluent and a chip; reagent standard curve information is stored in the chip; the sample diluent is used for diluting serum, plasma and/or whole blood samples to be detected.
The detection card of the invention utilizes fluorescent microsphere to mark, improves detection sensitivity, reduces non-specific combination, solves the problem of poor detection sensitivity of PCT/HBP, and has simple operation flow, short detection time and lower detection cost. In addition, the preparation method of the kit is simple, can realize batch production, is favorable for large-scale popularization and application, and has wide market prospect.
When the kit is used for detecting the content of procalcitonin and heparin binding protein in human serum, plasma and/or whole blood samples, the detection steps are as follows according to the reaction parameters shown in table 1:
firstly, adding a blood sample such as human serum, plasma and/or whole blood sample into a sample diluent, and uniformly mixing to obtain a mixed sample;
and secondly, adding the mixed sample to a sample pad of the test strip through a sample adding hole on a card shell of the detection card, namely, adding the mixed sample into the sample adding hole of the detection card, and displaying a detection result by an observation window on the card shell of the detection card within 10-15 minutes.
Finally, the intensity of fluorescent signals on the detection line of the reaction membrane reflects the quantity of the captured PCT/HBP antigens, and the concentration of the PCT/HBP antigens in the blood sample can be detected after the detection card is inserted into a matched immunofluorescence analyzer.
TABLE 1 reaction parameters
| Sample type | Sample size | Sample addition amount | Reaction time | Reaction temperature |
| Serum, plasma and whole blood | 100μL | 100μL | 15min | 18～30℃ |
The detection principle is as follows:
the kit adopts a double-antibody sandwich method and utilizes the technical principle of fluorescence immunochromatography. When in testing, the sample is evenly mixed with buffer solution and is dripped into a sample adding hole of the reagent, chromatography is carried out under the capillary effect, PCT/HBP antigen in the sample is combined with fluorescence labeling PCT/HBP antibody, and the mixture is diffused to a reaction membrane of a testing area and is captured by PCT/HBP monoclonal antibody coated by a detection line, and an antibody-antigen-fluorescence antibody complex is formed; the PCT/HBP concentration in the sample is proportional to the fluorescence intensity of the complex, and the immunofluorescence detector converts the fluorescence signal value into the PCT/HBP concentration in the sample according to a set standard curve.
The following is a further detailed description of the embodiments
Example 1 the composition, package and number (25 parts/box) of a procalcitonin/heparin binding protein immunochromatographic assay kit are shown in Table 2.
Table 2 composition, package and amount of kit
Example 2 test strip manufacturing Process
1) Preparation of sample pad
a) The sample pad is a glass fiber membrane, the sample pad is pretreated, soaked in boric acid-borax buffer solution containing BSA and Tween-20,0.2M with pH of 8.0+/-0.05, and dried for overnight (16-24) under the conditions of temperature (18-26) ℃ and relative humidity of less than or equal to 30 percent.
2) Preparation of bond pads
And (3) fully and uniformly mixing the prepared fluorescent microsphere marked PCT, HBP antibody, fluorescent microsphere marked procalcitonin/heparin binding protein antigen and fluorescent microsphere marked goat anti-chicken IgY antibody, uniformly spraying the mixture on the pretreated glass cellulose film at the concentration of 4 mu L/cm by using a quantitative film spraying instrument, and drying the mixture in an environment with the humidity of less than or equal to 30% at 37+/-2 ℃ for 16-24 hours. 3) Preparation of quality control line and detection line
The chicken IgY antibody and PCT/HBP (procalcitonin/heparin binding protein) antigen are diluted to the concentration of 1mg/ml respectively by using a coating liquid, namely 0.01M,0.01M pH7.4 PBS buffer containing sucrose, the distance between a quality control line and a detection line is 5+/-0.5 mm, the coating liquid is uniformly sprayed on a nitrocellulose membrane at the concentration of 1.0 mu L/cm by using a quantitative film spraying instrument, and the nitrocellulose membrane is placed in an electrothermal blowing drying box with the temperature of 37+/-2 ℃ and the humidity of less than or equal to 30 percent for drying for 16-24 hours.
4) Assembly of test strips
a) And assembling the test strip under the conditions that the humidity is less than 35 percent and the temperature is 20-30 ℃. The sample pad, the bonding pad, the reaction membrane and the absorption pad are sequentially connected in the direction of the adhesive surface of the bottom plate and the adjacent parts are partially overlapped.
b) When the NC film is adhered to the bottom plate, the position of the detection line is close to the bonding pad, the position of the quality control line is close to the absorption pad, and the assembled NC film is cut into test strips with the width of 3.85+/-0.05 mm.
c) And the test paper strip is correctly arranged on the bottom plate, the absorption pad is arranged at the terminal of the bottom plate, and the absorption pad is arranged on the shell pressing equipment to be firmly pressed to obtain the test paper card for later use.
Example 3ID chip entry of stored data information
And taking test strips and buffer solution which are qualified in intermediate product detection, wherein each calibrator has at least 5 concentrations, each concentration is tested for at least 3 times, and the average value of T/C values is obtained through calculation and test.
And taking the prepared concentration and the corresponding T/C average value as linear data, substituting the data into special software to fire the chip, and sticking a chip label.
It is to be understood that the foregoing description of the preferred embodiments of the invention is not to be considered as limiting the scope of the invention, which is defined by the appended claims.
Claims (10)
1. A dry fluorescent immunochromatographic assay card for quantitatively detecting procalcitonin and heparin binding proteins, which is characterized by comprising a test strip and a card shell adapted to accommodate the test strip; wherein:
the test strip comprises a sample pad, a combination pad, a reaction membrane, an absorption pad and a bottom plate; the bottom plate is in a strip structure, the reaction film is stuck to the middle part of one surface of the bottom plate, and the left side and the right side of the reaction film are respectively laminated, buckled and lapped with one side of the bonding pad and one side of the absorption pad; one side edge of the sample pad is laminated, buckled and overlapped on the other side edge of the bonding pad, and the sample pad and the absorption pad are respectively arranged at two ends of the bottom plate; the binding pad is coated with procalcitonin/heparin binding protein marked by fluorescent microspheres and sheep anti-chicken IgY antibody marked by fluorescent microspheres; two detection lines and a quality control line which are parallel to each other are arranged in the middle of the reaction film, and the two detection lines are adjacent; the detection line is coated with procalcitonin/heparin binding protein antigen; the quality control line is coated with chicken IgY monoclonal antibody;
the clamping shell comprises a containing cavity, a sample adding hole and an observation window; the sample adding hole and the observation window are respectively communicated with the accommodating cavity; when the test strip is placed in the accommodating cavity of the cartridge, the sample pad is exposed through the sample adding hole, and the reaction membrane is exposed through the observation window.
2. The dry fluorescent immunochromatographic assay card according to claim 1, wherein the binding pad is coated with a fluorescent microsphere-labeled procalcitonin/heparin binding protein antibody recognizing different epitopes and a fluorescent microsphere-labeled goat anti-chicken IgY monoclonal antibody.
3. The dry fluorescent immunochromatographic assay card according to claim 1, wherein the detection line is spaced apart from the quality control line by a distance of 5.+ -. 0.5mm.
4. The dry fluorescent immunochromatographic assay card according to claim 1, wherein the coating liquid for coating the procalcitonin/heparin binding protein antigen and the chicken IgY monoclonal antibody is a mixed liquid comprising 0.01M sucrose and 0.01M PBS buffer with pH of 7.4.
5. The dry fluorescent immunochromatographic assay card according to claim 4, wherein the concentration of the mixed solution is 1mg/ml.
6. The dry fluorescent immunochromatographic assay card according to claim 1, wherein the sample pad is a glass cellulose membrane dried after being immersed in a surfactant buffer.
7. The dry fluorescent immunochromatographic assay card according to claim 6, wherein the active agent buffer is a Tween-20 containing BSA, pH 8.0.+ -. 0.05 and boric acid-borax buffer of 0.2M.
8. The dry-type fluorescence immunochromatographic assay card according to claim 1, wherein the reaction membrane is made of a nitrocellulose membrane; the bonding pad is made of a glass cellulose film.
9. A dry fluorescent immunochromatographic kit for quantitatively detecting procalcitonin and heparin binding proteins, which is characterized by comprising the dry fluorescent immunochromatographic detection card according to any one of claims 1 to 8, a sample diluent and a chip; reagent standard curve information is stored in the chip; the sample diluent is used for diluting serum, plasma and/or whole blood samples to be detected.
10. A method for detecting procalcitonin and heparin binding protein levels in serum, plasma and/or whole blood using the kit of claim 9, comprising the steps of:
firstly, adding serum, plasma and/or whole blood samples into a sample diluent, and uniformly mixing to obtain a mixed sample;
secondly, adding the mixed sample to a sample pad of the test strip, and displaying a detection result by a detection card within 10-15 minutes;
finally, after the detection card in the kit is inserted into a matched immunofluorescence analyzer, the concentration of procalcitonin and heparin binding protein in serum, plasma and/or whole blood samples is detected.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210033632.2A CN116466072A (en) | 2022-01-12 | 2022-01-12 | Dry-type fluorescence immunochromatography detection card, kit and detection method for quantitatively detecting procalcitonin and heparin binding protein |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210033632.2A CN116466072A (en) | 2022-01-12 | 2022-01-12 | Dry-type fluorescence immunochromatography detection card, kit and detection method for quantitatively detecting procalcitonin and heparin binding protein |
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