CN105606825A - Fluorescence immunochromatography detection kit for gastrin releasing peptide precursor - Google Patents
Fluorescence immunochromatography detection kit for gastrin releasing peptide precursor Download PDFInfo
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- CN105606825A CN105606825A CN201610068561.4A CN201610068561A CN105606825A CN 105606825 A CN105606825 A CN 105606825A CN 201610068561 A CN201610068561 A CN 201610068561A CN 105606825 A CN105606825 A CN 105606825A
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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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- 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
- G01N33/57407—Specifically defined cancers
- G01N33/57423—Specifically defined cancers of lung
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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
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
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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
-
- 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
- G01N2333/5758—Gastrin releasing peptide
Abstract
The invention belongs to the field of medical detection and specifically relates to a fluorescence immunochromatography detection kit for a gastrin releasing peptide precursor. The fluorescence immunochromatography detection kit comprises a test paper card and a fluorescence immunochromatography analysis meter. In an embodiment of the invention, the test paper card is formed according to the following steps: mutually overlapping a treated sample pad, a combining pad adsorbing rare earth fluorescent microsphere labelled antibody, a nitrocellulose membrane coated with a detecting line and a quality control line and a water absorbing pad on a PVC (Poly Vinyl Chloride) plate in turn, assembling into test paper, cutting the test paper into 4mm wide test paper strips and storing the test paper strips in a plastic card, thereby forming the test paper card.
Description
Technical field
The invention belongs to medical science detection field, be specifically related to a kind of gastrin-releasing peptide precursor fluorescence immune chromatography detection kit.
Background technology
Lung cancer is one of the incidence of disease and the highest malignant tumour of case fatality rate in the world. Lung cancer is divided into non-small cell lung cancer (NSCLC) and ED-SCLC (SCLC), and the latter accounts for the 15%-20% of patients with lung cancer sum. SCLC is a kind of malignant tumour of Fast Growth, and patient's majority when medical has been found that regional nodes or DISTANT METASTASES IN, and early diagnosis can improve survival rate. It is an important aided detection method of lung cancer that tumor markers detects, desirable tumor markers should have higher sensitivity and specificity, not only contribute to the early diagnosis of tumour, judgement and the cancer staging of histological type, and can assess result for the treatment of and prognosis and guides individualized treatment. At present conventional lung cancer related neoplasms mark carcinomebryonic antigen (CEA), the soluble fragments (CYFRA21-1) of Cyfra21-1, squamous cell carcinoma antigen (SCCA), neuronspecific enolase (NSE) are not high to early stage of lung cancer diagnostic sensitivity and specificity, are not suitable as early stage of lung cancer examination. Studies show that in a large number, tumor markers gastrin-releasing peptide precursor (proGRP) has higher clinical value.
People's gastrin-releasing peptide precursor (proGRP) is newfound a kind of tumor markers for SCLC in recent years, and it not only can be used for the early diagnosis of SCLC, also contributes to judge curative effect and early detection tumor recurrence. The critical value of gastrin-releasing peptide precursor in human serum is 50pg/ml, be 100% to the specificity of SCLC early diagnosis, detection sensitivity is 75% left and right, and the specificity of its diagnosis SCLC can reach more than 95%, and therefore gastrin-releasing peptide precursor is the good index of ED-SCLC auxiliary diagnosis.
Gastrin-releasing peptide precursor detection method mainly contains double antibodies sandwich method (ELISA), immunoluminescence method, radio immunoassay. Wherein, ELISA standard measure poor accuracy, the operating time is long, automaticity is low, is used for qualitative detection; Radio immunoassay sensitivity can reach 4pg/ml, can detect normal human serum gastrin-releasing peptide precursor, sensitiveer than double antibodies sandwich method, and shortcoming is that required time is longer, and testing result is unstable, and repeatability is poorer than ELISA, and has radioactive pollution danger; Immunoluminescence method method high specificity, sensitiveness is high, but needs expensive instrument and equipment and veteran operating personnel, general how use in specific medical mechanism. Therefore develop that sensitivity is higher, gastrin-releasing peptide precursor product fast and easily, be still clinical diagnosis product research field and need badly the major issue of solution.
Summary of the invention
The object of this invention is to provide a kind of gastrin-releasing peptide precursor quantitative fluorescence immunochromatographytest test kit; It is made up of test card and fluorescence immune chromatography analyzer.
In one embodiment of the invention, described test card is that the treated sample pad of overlap joint, absorption have the pad of rare-earth fluorescent microballoon labelled antibody, the nitrocellulose filter that is coated with detection line and nature controlling line and adsorptive pads mutually in turn on PVC plate, after assembling, form test paper, then cut into the wide test strips of 4mm, test strips is packed into and in plastic clip, forms test card.
In the further embodiment of the present invention, described rare-earth fluorescent microballoon, doped with lanthanide series, comprises any one or a few of the lanthanide series such as europium (Eu), samarium (Sm), erbium (Er) and/or neodymium (Nd). The diameter of described rare-earth fluorescent microballoon is 60-200nm, and further preferably, the diameter of selected rare-earth fluorescent microballoon is 100-120nm.
In the present invention, described rare-earth fluorescent microballoon contains rare-earth complex, stable under ground state, under the excitation source effect of 350-400nm, can launch the fluorescence of wave-length coverage at 550-600nm.
Described fluorescence immune chromatography analyzer is a kind of Systems for optical inspection, for the quantitative judgement to testing result, is 0-5ng/mL to the detection range of gastrin-releasing peptide precursor.
In another embodiment of the invention, the pad of the antibody of described absorption fluorescent microsphere mark is prepared by the following method:
The aldehyde radical of A, rare-earth fluorescent microballoon:
Get 5mg rare-earth fluorescent microballoon, with 50mM, the carbonate buffer solution of pH9.5, adopts centrifugal process washing 3 times, and centrifugal speed is 12000rpm, and the time is 10 minutes, is finally resuspended in the above-mentioned carbonate buffer solution of 200 μ l. The glucan that adds 400 μ l aldehyde radicals, mixes, dark reaction 4 hours under room temperature. Adopt the washing of same centrifugal process and be resuspended in the above-mentioned carbonate buffer solution of 200 μ l, be placed in 4 DEG C for subsequent use.
The preparation of the gastrin-releasing peptide precursor monoclonal antibody of B, rare-earth fluorescent microballoon mark:
By 2mg gastrin-releasing peptide precursor monoclonal antibody with above-mentioned carbonate buffer solution in 4 DEG C of dialysed overnight, then mix with the rare-earth fluorescent microballoon of above-mentioned aldehyde radical, 4 DEG C of reactions are spent the night. Then, add sodium borohydride to final concentration 10mM, 4 DEG C are reacted 4 hours; Add isopyknic confining liquid (100mM TRIS buffer, pH8.0 contain 0.5%BSA, 0.1% sucrose and 0.1%PVP), 4 DEG C of sealings are spent the night again; Then use 100mM TRIS buffer, the buffer solution of pH8.0 adopts centrifugal process washing 3 times, is resuspended in the 100mM TRIS buffer of 200 μ l (containing 1.0%NaCl, 0.3%BSA, 0.2% ethylene glycol), and 4 DEG C keep in Dark Place for subsequent use.
The preparation of the pad of the antibody of C, absorption fluorescent microsphere mark:
Glass fibre membrane is soaked in to (containing 1.2%TritonX-100,2.5%BSA, pH8.0) in 250mM TRIS buffer, and 4 DEG C are soaked 4 hours, then take out 37 DEG C of oven for drying 4 hours, for subsequent use. By glass fibre membrane on the three-dimensional specking platform of Bio-DotXYZ3050, the gastrin-releasing peptide precursor monoclonal antibody of rare-earth fluorescent microballoon mark is sprayed onto to glass fibre membrane with the Bio-JetQuanti300 noncontact quantitation nozzle that declines, dry 4 hours for 37 DEG C, for subsequent use.
In another embodiment of the present invention, described in to be coated with the preparation method of nitrocellulose filter of detection line and nature controlling line as follows:
Gastrin-releasing peptide precursor monoclonal antibody and goat anti-mouse igg antibody are adjusted to concentration to 5mg/ml with coated dilution, film liquid measure is 2 μ l/cm, they are coated with as be sprayed on nitrocellulose filter parallel with nature controlling line of detection line respectively, detection line and nature controlling line are spaced apart 5mm, then be placed in baking oven, dry 4 hours for 37 DEG C. Described coated dilution is 100mM TRIS buffer, pH8.0.
In another embodiment of the present invention, the preparation method of described sample pad is as follows:
Glass fibre membrane is soaked in to 0.25MTris buffer solution (pH7.5), and 1.2%TritonX-100, in the treatment fluid of 2.5%BSA, in 4 DEG C of immersions 4 hours, is then placed in baking oven, dries 4 hours for 37 DEG C.
In the present invention, the assembling process of described test card is as follows: on PVC plate, pasting successively treated sample pad, absorption has pad, the nitrocellulose filter that is coated with detection line and nature controlling line and the adsorptive pads of antibody of rare-earth fluorescent microballoon mark, after assembling, obtain the large plate of test paper, cut into as requested 4mm wide, test paper is packed into and in plastic clip, forms test card.
Described kit, detection method is: 1) will detect reagent and sample balance to room temperature, and take out test card, and keep flat; 2) accurately draw 100 μ l serum samples, when sample is whole blood, draw 150 μ l samples, join in sample aperture, in 15-30 minute, quantitatively judge result with fluorescence immune chromatography analyzer; 3) set and test card is put into storehouse after instrument relevant parameter and detect, instrument will demonstrate the quantitative assay result of sample concentration.
Detailed description of the invention
To describe the present invention below in detail. It is pointed out that following explanation is only illustrating of the technical scheme claimed to the present invention, the not any restriction to these technical schemes. The content that protection scope of the present invention is recorded with appended claims is as the criterion.
The preparation of embodiment 1 gastrin-releasing peptide precursor quantitative fluorescence immunochromatographytest test kit
1) preparation of the pad of the antibody of absorption fluorescent microsphere mark:
The aldehyde radical of A, rare-earth fluorescent microballoon:
Get 5mg rare-earth fluorescent microballoon (diameter 100nm), with 50mM, the carbonate buffer solution of pH9.5, adopts centrifugal process washing 3 times, and centrifugal speed is 12000rpm, and the time is 10 minutes, is finally resuspended in the above-mentioned carbonate buffer solution of 200 μ l. The glucan that adds 400 μ l aldehyde radicals, mixes, dark reaction 4 hours under room temperature. Adopt the washing of same centrifugal process and be resuspended in the above-mentioned carbonate buffer solution of 200 μ l, be placed in 4 DEG C for subsequent use.
The preparation of the gastrin-releasing peptide precursor monoclonal antibody of B, rare-earth fluorescent microballoon mark:
By 2mg gastrin-releasing peptide precursor monoclonal antibody with above-mentioned carbonate buffer solution in 4 DEG C of dialysed overnight, then mix with the rare-earth fluorescent microballoon of above-mentioned aldehyde radical, 4 DEG C of reactions are spent the night. Then, add sodium borohydride to final concentration 10mM, 4 DEG C are reacted 4 hours; Add isopyknic confining liquid (100mM TRIS buffer, pH8.0 contain 0.5%BSA, 0.1% sucrose and 0.1%PVP), 4 DEG C of sealings are spent the night again; Then use 100mM TRIS buffer, the buffer solution of pH8.0 adopts centrifugal process washing 3 times, is resuspended in the 100mM TRIS buffer of 200 μ l (containing 1.0%NaCl, 0.3%BSA, 0.2% ethylene glycol), and 4 DEG C keep in Dark Place for subsequent use.
The preparation of the pad of the antibody of C, absorption fluorescent microsphere mark:
Glass fibre membrane is soaked in to (containing 1.2%TritonX-100,2.5%BSA, pH8.0) in 250mM TRIS buffer, and 4 DEG C are soaked 4 hours, then take out 37 DEG C of oven for drying 4 hours, for subsequent use. By glass fibre membrane on the three-dimensional specking platform of Bio-DotXYZ3050, the gastrin-releasing peptide precursor monoclonal antibody of rare-earth fluorescent microballoon mark is sprayed onto to glass fibre membrane with the Bio-JetQuanti300 noncontact quantitation nozzle that declines, dry 4 hours for 37 DEG C, for subsequent use.
2) be coated with the preparation of the nitrocellulose filter of detection line and nature controlling line:
Gastrin-releasing peptide precursor monoclonal antibody and goat anti-mouse igg antibody are adjusted to concentration to 5mg/ml with coated dilution, film liquid measure is 2 μ l/cm, they are coated with as be sprayed on nitrocellulose filter parallel with nature controlling line of detection line respectively, detection line and nature controlling line are spaced apart 5mm, then be placed in baking oven, dry 4 hours for 37 DEG C. Described coated dilution is 100mM TRIS buffer, pH8.0.
3) preparation of sample pad:
Glass fibre membrane is soaked in to 0.25MTris buffer solution (pH7.5), and 1.2%TritonX-100, in the treatment fluid of 2.5%BSA, in 4 DEG C of immersions 4 hours, is then placed in baking oven, dries 4 hours for 37 DEG C.
4) assembling process is as follows: on PVC plate, pasting successively treated sample pad, absorption has pad, the nitrocellulose filter that is coated with detection line and nature controlling line and the adsorptive pads of antibody of rare-earth fluorescent microballoon mark, after assembling, obtain the large plate of test paper, cut into as requested 4mm wide, test paper is packed into and in plastic clip, forms test card.
In test card, the monoclonal antibody in the pad of the antibody of absorption fluorescent microsphere mark and be coated with monoclonal antibody in the nitrocellulose filter of detection line and nature controlling line for pairing antibody, purchased from quark bio tech ltd, Zhejiang.
The foundation of embodiment 2 kit calibration curves
Get 0,0.05,0.1,0.5,1, the gastrin-releasing peptide precursor standard items of 5ng/ml, measure with test card,
1) will detect reagent and sample balance to room temperature, take out test card (prepared by embodiment 1), keep flat; 2) accurately draw 100 μ l standard items samples, join in the sample aperture of test card, after 20 minutes, detect with fluorescence immune chromatography analyzer.
With fluorescence intensity level and corresponding standard items concentration Criterion curve, be specially Y(concentration of specimens)=0.067X+0.012, R2Value > 0.99, illustrate that kit of the present invention is linear good.
Embodiment 3 kit study on the stability
Kit prepared by embodiment 1 is placed 3 months under 37 DEG C of environment, then measures calibration curve and the R of kit according to the method for embodiment 22Value, and adopt the gastrin-releasing peptide precursor standard items of 1ng/ml to carry out precision investigation (CV% is calculated in parallel determination 10 times) to kit, concrete outcome is as follows:
Comparative example 1 preparation method is with embodiment 1, difference is only that the confining liquid in the preparation of gastrin-releasing peptide precursor monoclonal antibody of rare-earth fluorescent microballoon mark exists different, the confining liquid of comparative example 1 consists of: contain 0.5%BSA, the 100mMTris buffer solution of 0.1% sucrose and 0.1%PVP, pH7.5.
Comparative example 2 preparation methods are with embodiment 1, difference is only that the confining liquid in the preparation of gastrin-releasing peptide precursor monoclonal antibody of rare-earth fluorescent microballoon mark exists different, the confining liquid of comparative example 2 consists of: the 100mM TRIS buffer buffer solution that contains 0.5%BSA and 0.1% sucrose, pH8.0.
Comparative example 3 preparation methods are with embodiment 1, difference is only that the confining liquid in the preparation of gastrin-releasing peptide precursor monoclonal antibody of rare-earth fluorescent microballoon mark exists different, the confining liquid of comparative example 3 consists of: the 100mM TRIS buffer buffer solution that contains 2%BSA and 5% sucrose, pH8.0.
Comparative example 4 preparation methods are with embodiment 1, difference is only that the confining liquid in the preparation of gastrin-releasing peptide precursor monoclonal antibody of rare-earth fluorescent microballoon mark exists different, the confining liquid of comparative example 4 consists of: contain 0.5%BSA, the 100mM TRIS buffer buffer solution of 0.1% sucrose and 0.1%PVP, pH7.5.
Comparative example 5 preparation methods are with embodiment 1, difference is only that the resuspended liquid in the preparation of gastrin-releasing peptide precursor monoclonal antibody of rare-earth fluorescent microballoon mark exists different, the resuspended liquid of comparative example 5 consists of: the 100mM TRIS buffer buffer solution that contains 1.0%NaCl and 0.3%BSA, pH8.0.
Content of the present invention only for example understands some claimed specific embodiments; the technical characterictic of recording in one of them or more technical scheme can be with one or more technical schemes be combined arbitrarily; these technical schemes that obtain through combination are also in the application's protection domain, just as these technical schemes that obtain through combination have specifically been recorded in the disclosure of invention.
Claims (6)
1. a gastrin-releasing peptide precursor quantitative fluorescence immunochromatographytest test kit; It is made up of test card and fluorescence immune chromatography analyzer; Described test card is that the treated sample pad of overlap joint, absorption have the pad of rare-earth fluorescent microballoon labelled antibody, the nitrocellulose filter that is coated with detection line and nature controlling line and adsorptive pads mutually in turn on PVC plate, after assembling, form test paper, then cut into the wide test strips of 4mm, test strips is packed into and in plastic clip, forms test card.
2. immunochromatographytest test kit according to claim 1, it is characterized in that, described rare-earth fluorescent microballoon, doped with lanthanide series, comprises any one or a few of the lanthanide series such as europium (Eu), samarium (Sm), erbium (Er) and/or neodymium (Nd); The diameter of described rare-earth fluorescent microballoon is 60-200nm.
3. immunochromatographytest test kit according to claim 2, is characterized in that, the diameter of selected rare-earth fluorescent microballoon is 100-120nm.
4. immunochromatographytest test kit according to claim 1, is characterized in that, the pad of the antibody of described absorption fluorescent microsphere mark is prepared by the following method:
The aldehyde radical of A, rare-earth fluorescent microballoon:
Get 5mg rare-earth fluorescent microballoon, with 50mM, the carbonate buffer solution of pH9.5, adopts centrifugal process washing 3 times, and centrifugal speed is 12000rpm, and the time is 10 minutes, is finally resuspended in the above-mentioned carbonate buffer solution of 200 μ l; The glucan that adds 400 μ l aldehyde radicals, mixes, dark reaction 4 hours under room temperature; Adopt the washing of same centrifugal process and be resuspended in the above-mentioned carbonate buffer solution of 200 μ l, be placed in 4 DEG C for subsequent use;
The preparation of the gastrin-releasing peptide precursor monoclonal antibody of B, rare-earth fluorescent microballoon mark:
By 2mg gastrin-releasing peptide precursor monoclonal antibody with above-mentioned carbonate buffer solution in 4 DEG C of dialysed overnight, then mix with the rare-earth fluorescent microballoon of above-mentioned aldehyde radical, 4 DEG C of reactions are spent the night; Then, add sodium borohydride to final concentration 10mM, 4 DEG C are reacted 4 hours; Add isopyknic confining liquid (100mM TRIS buffer, pH8.0 contain 0.5%BSA, 0.1% sucrose and 0.1%PVP), 4 DEG C of sealings are spent the night again; Then use 100mM TRIS buffer, the buffer solution of pH8.0 adopts centrifugal process washing 3 times, is resuspended in the 100mM TRIS buffer of 200 μ l (containing 1.0%NaCl, 0.3%BSA, 0.2% ethylene glycol), and 4 DEG C keep in Dark Place for subsequent use;
The preparation of the pad of the antibody of C, absorption fluorescent microsphere mark:
Glass fibre membrane is soaked in to (containing 1.2%TritonX-100,2.5%BSA, pH8.0) in 250mM TRIS buffer, and 4 DEG C are soaked 4 hours, then take out 37 DEG C of oven for drying 4 hours, for subsequent use; By glass fibre membrane on the three-dimensional specking platform of Bio-DotXYZ3050, the gastrin-releasing peptide precursor monoclonal antibody of rare-earth fluorescent microballoon mark is sprayed onto to glass fibre membrane with the Bio-JetQuanti300 noncontact quantitation nozzle that declines, dry 4 hours for 37 DEG C, for subsequent use.
5. immunochromatographytest test kit according to claim 1, is characterized in that, described in to be coated with the preparation method of nitrocellulose filter of detection line and nature controlling line as follows:
Gastrin-releasing peptide precursor monoclonal antibody and goat anti-mouse igg antibody are adjusted to concentration to 5mg/ml with coated dilution, film liquid measure is 2 μ l/cm, they are coated with as be sprayed on nitrocellulose filter parallel with nature controlling line of detection line respectively, detection line and nature controlling line are spaced apart 5mm, then be placed in baking oven, dry 4 hours for 37 DEG C; Described coated dilution is 100mM TRIS buffer, pH8.0.
6. immunochromatographytest test kit according to claim 1, is characterized in that, the preparation method of described sample pad is as follows:
Glass fibre membrane is soaked in to 0.25MTris buffer solution (pH7.5), and 1.2%TritonX-100, in the treatment fluid of 2.5%BSA, in 4 DEG C of immersions 4 hours, is then placed in baking oven, dries 4 hours for 37 DEG C.
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| CN107389928A (en) * | 2017-08-31 | 2017-11-24 | 重庆康巨全弘生物科技有限公司 | Quantitatively detect two-photon fluorescence immune chromatography reagent kit of gastrin-releasing peptide precursor (pro GRP) and preparation method thereof |
| CN108931636A (en) * | 2018-07-02 | 2018-12-04 | 威海纽普生物技术有限公司 | Measure the fluorescence immune chromatography detection kit and preparation method thereof of ST2 |
| CN109324181A (en) * | 2018-10-22 | 2019-02-12 | 河北特温特生物科技发展有限公司 | A kind of preparation method of immunochromatography sealant compositions, purposes and immune chromatography reagent kit |
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