CN103913568A - Joint inspection test paper strip for human body autoantibody and preparation method thereof - Google Patents
Joint inspection test paper strip for human body autoantibody and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a joint inspection test paper strip for human body autoantibody and a preparation method thereof. The the test strip comprises a sample pad, a glass fiber membrane containing a colloidal gold particle marker, a nitrocellulose membrane and absorbent paper, which are in successive overlap joint. The nitrocellulose membrane includes a detection area coated with systemic lupus erythematosus (sm) recombinant antigen and systemic sclerosis (Scl-70) and a control area coated with sheep antimice antibody; the colloidal gold particle marker comprises a micro signal amplification system and colloidal gold marked mice IgG, and the micro signal amplification system is colloidal gold particle-avidin-biotin-sm/Scl-70 recombinant antigen. The joint inspection test paper strip is obtained by adding the biotin-avidin micro signal amplification system in a double antigen sandwich detection system, a signal of a target antibody is expanded, the detection sensitivity is increased, false negative or leak detection caused by a too weak signal can be avoided, at the same time, synchronous joint inspection of systemic lupus erythematosus (sle) and systemic sclerosis can be performed, and the test time, test sample and cost are saved.
Description
Technical field
The invention belongs to field of medical examination, relate in particular to a kind of human body autoantibody joint inspection test strips and preparation method thereof.
Background technology
The antibody of autoantibody pointer to autologous tissue, organ, cell and cell component.Growth, growth and the existence of human body has maintaining of complete autoimmune tolerance mechanism, and normal immune response has protectiveness defense reaction, autologous tissue, composition is not reacted.Once the integrality of self tolerance is destroyed, body is looked autologous tissue, composition is " foreign matter ", and autoimmune response occurs, and produces autoantibody.These autoantibodies can be attacked body self cell, tissue, organ, and the reaction that causes inflammation, damages body.It is sclerosis, neuromuscular disease, multiple sclerosis, myasthenia gravis, demyelinating disease, incretion disease, primary adrenal cortical atrophy, chronic first shape inflammation, juvenile onset diabetes, disease of digestive system chronic nonspecific ulcerative colitis, chronic active hepatitis, bad habit anaemia and atrophic gastritis etc. that common autoimmune disease includes connective tissue disease, rheumatoid arthritis, systemic loupus erythematosus, dermatomyositis, system.
Wherein systemic loupus erythematosus and systemic sclerosis in connective tissue disease (CTD), rank first place respectively position and second.Systemic loupus erythematosus (systemic lupus erythematosus, SLE) be a kind of diffusivity, general autoimmunity disease, mainly involve mucocutaneous, skeletal muscle, kidney and central nervous system, also can involve multiple organs and the systems such as lung, heart, blood simultaneously, occur various clinical symptoms; In serum, multiple autoantibody and crucial immunological abnormality can be detected.SLE is apt to occur in young women, and onset peak is 15～40 years old, and men and women's morbidity ratio is 1: 9 left and right.The men and women's of childhood and senile SLE ratio is about 1: 2.The morbidity rate in the whole world is about 30～50,/10 ten thousand people, and the morbidity rate of China is about 70,/10 ten thousand people.Although the morbidity of SLE is subject to the impact of inherent cause, great majority are Sporadic cases.Systemic sclerosis (chorionitis) is a kind of connective tissue disease that is hardened to feature with the each system collagenous fibres of skin.The systemic scleroderma of involving internal organs claims again systemic sclerosis.This disease is only second to lupus erythematosus and occupies second in connective tissue disease (CTD).Patient is more with women, and women is about 3: 1 with the male sex's ratio.Age of onset is common with 20～50 years old.
Sm antibody normal reference value is negative, is the serum marker antibody of systemic loupus erythematosus (SLE) clinically.Scl-70 antibody normal reference value is negative, is the serum marker antibody of diffuse systemic sclerosis (PSS) clinically.
As the important blood serum designated object of autoimmunity disease, the fast detecting of human body autoantibody has important using value and theory significance for diagnosis, the prognosis of autoimmune disease and the pathologic process that lapses to judgement and understand disease.
Sm antibody and Scl-70 antibody all belong to soluble antigen antibody (ENA) category in anti-core, and conventional detection method has gel countercurrent electrophoresis, immune double diffusion and Western blot.At present the most frequently used is Western blot, adopt Western blotting law technology, by gel electrophoresis, hybrid antigen is carried out to protein molecular separation, again the protein molecular of separating is transferred on film, film is dried, cut into inch strips, be made into and detect film bar, then be used in conjunction with other constituents of kit.Can disposablely detect multiple autoantibody disease and the advantage such as cheap although the method has, manufacture craft is comparatively numerous and diverse simultaneously, and length consuming time is unfavorable for popularizing, and background colour complexity, is unfavorable for the interpretation of result.
As mentioned above, based on nm of gold detection technique, add again Avidin-Biotin signal and expand system, in conjunction with recombinant antigen technology of preparing, simultaneously by adjusting different preparation technology parameter, overcoming nm of gold under-sensitive and also can realize the synchronous detection of systemic loupus erythematosus and systemic sclerosis simultaneously.
Summary of the invention
The object of the invention is to solve the autoantibody detection film bar manufacture craft existing in prior art numerous and diverse, length consuming time, is unfavorable for popularizing, and background colour complexity, be unfavorable for the defects such as the interpretation of result, a kind of human body autoantibody joint inspection test strips and preparation method thereof is provided.
For achieving the above object, the present invention has taked following technical scheme:
1. a human body autoantibody joint inspection test strips, comprise sample pad (1), the glass fibre membrane that contains colloid gold particle label (2) being closely connected with described sample pad (1) one end, the nitrocellulose filter (3) being closely connected with described glass fibre membrane (2) other end, and the thieving paper (4) being closely connected with the other end of described nitrocellulose filter (3), described sample pad (1), glass fibre membrane (2), nitrocellulose filter (3) and thieving paper (4) are all arranged on base plate (5), described nitrocellulose filter (3) is coated with the detection zone (6) of systemic loupus erythematosus sm recombinant antigen and the detection zone (7) of systemic sclerosis Scl-70 recombinant antigen, and be coated with the control zone (8) of sheep anti-mouse antibody, described colloid gold particle label comprises the mouse IgG of micro-signal amplification system and colloid gold label, described micro-signal amplification system is colloid gold particle-Avidin-Biotin-sm/Scl-70 recombinant antigen.
Preferably, described systemic loupus erythematosus sm recombinant antigen consumption is 0.13-0.27 μ g/mm, and described systemic sclerosis Scl-70 recombinant protein consumption is 0.13-0.27 μ g/mm, and described colloid gold label has mouse IgG antibody, and consumption is 15-50 μ g/cm
2.The consumption of described colloid gold particle-Avidin-Biotin-Sm/SCL-70 recombinant antigen compound is 12.5-68 μ g/cm
2.Described sheep anti-mouse antibody is sheep anti-mouse igg antibody, and the consumption of described sheep anti-mouse antibody is 0.20-0.48 μ g/mm.
The present invention also provides a kind of preparation method of above-mentioned test strips, comprises the following steps:
(1) preparation system lupus erythematosus sm recombinant antigen, systemic sclerosis Scl-70 recombinant antigen and colloid gold label mouse IgG antibody;
(2) prepare micro-signal amplification system
A, prepare colloid gold label Avidin
By Avidin to be marked dialysed overnight in the NaCl of 0.001～0.01mol/L solution in advance, centrifugal, pH value to 9～10 of adjusting collaurum liquid, mark Avidin is to form colloid gold label Avidin;
B, prepare biotinylation sm recombinant antigen and biotinylation Scl-70 recombinant antigen according to a conventional method;
C, complete micro-signal amplification system
The biotinylation sm/Scl-70 recombinant antigen that the colloid gold label Avidin that step a is obtained and step b obtain mixes, and connects and obtains collaurum-Avidin-Biotin-sm/Scl-70 recombinant antigen compound, and washing, obtains micro-signal amplification system;
(3) by dilution parameters 25～40cm
2/ ml, is sprayed onto the collaurum-Avidin-Biotin of step (2)-sm recombinant antigen and collaurum-Avidin-Biotin-Scl-70 recombinant antigen on glass fibre membrane, dry more than 12 hours, for subsequent use;
(4) with coated damping fluid dilution sm recombinant antigen, making its concentration is 1.0～1.5mg/ml, by film liquid measure 0.15～0.2 μ l/mm, by its careful being sprayed onto uniformly on nitrocellulose filter, dry more than 12 hours, for subsequent use;
With coated damping fluid dilution Scl-70 recombinant antigen, making its concentration is 1.0～1.5mg/ml, by film liquid measure 0.15～0.20 μ l/mm, by its careful being sprayed onto uniformly on nitrocellulose filter, dry more than 12 hours, for subsequent use;
With coated damping fluid dilution sheep anti-mouse antibody, making its concentration is 1.0～2.0mg/ml, by film liquid measure 0.16～0.20 μ l/mm, by its careful being sprayed onto uniformly on nitrocellulose filter, dry more than 12 hours, for subsequent use;
(5) glass fibre membrane of sample pad, step (3), the nitrocellulose filter of step (4), thieving paper are pasted on base plate according to the order of sequence, obtain final product.
Expand not enough and can not be by the problem of the synchronous human body autoantibody of immunochromatographic method disease in order to solve signal, the present invention is based on solid-phase immunity chromatographic theory, adopt systemic loupus erythematosus sm antibody and systemic sclerosis Scl-70 antibody in functional color micro-sphere technology (colloid gold particle labelling technique), micro-signal amplifying technique (Avidin-Biotin system and double antigens sandwich reaction system) and joint inspection technology for detection blood of human body.If contain systemic loupus erythematosus sm antibody or systemic sclerosis Scl-70 antibody in sample, micro-signal cascade system on glass fibre membrane is amplified signal, sm/Scl-70 antibody is combined with colloid gold particle label, form compound, and be diffused on nitrocellulose filter further chromatography, when running into while being coated on the pairing antigen that the detection line 6 of detection zone on nitrocellulose filter or detection line 7 (being respectively the coated line of systemic loupus erythematosus sm recombinant antigen or systemic sclerosis Scl-70 recombinant antigen) locate, compound again with envelope antigen combination, be trapped in coated place, in the time that captive compound reaches some, form macroscopic detection lines, in interpret sample, contain sm antibody or Scl-70 antibody, if do not occur, negative or the content of interpret sample is lower than the lowest detectable limit of test strips.Control zone (C line) is as the quality control standard of test strips, and positive and negative sample all there will be while detection.
Compared with prior art, the present invention has following beneficial effect:
(1) the present invention, on the basis of existing test strip, has added biotin-avidin micro-signal cascade amplification system, thereby, in the process of detection autoantibody, expand the signal of target antibody, increased detection sensitivity, avoided occurring because signal is too weak false cloudy or undetected;
(2) test strips of the present invention there is handling safety (polluting without radiation), easy (simple operations one step completes), be applicable to single/part detect (that enzyme exempts to be not suitable for is single/part or a small amount of sample detection) and advantage such as (can have result about 3-5 minute) fast, can realize systemic loupus erythematosus and system is the POCT detection of sclerosis;
(3) test strips of the present invention can realize while detection system lupus erythematosus, systemic sclerosis on a paper slip, has shortened detection time with respect to western blotting method; With respect to market existing autoimmune disease immuno-chromatographic test paper strip testing product, increase detection sensitivity, and realized the synchronous detection of 2 kinds of autoimmune diseases, save medical disposable material.
Accompanying drawing explanation
Fig. 1 is the structural representation of a kind of human body autoantibody joint inspection test strips of the present invention;
Fig. 2 is the effective assay schematic diagram that utilizes a kind of human body autoantibody of the present invention joint inspection ELISA test strip systemic loupus erythematosus and systemic sclerosis
Fig. 3 is the null result assay schematic diagram that utilizes a kind of human body autoantibody of the present invention joint inspection ELISA test strip systemic loupus erythematosus and systemic sclerosis
Reference numeral: 1. sample pad; 2. glass fibre membrane; 3. nitrocellulose filter; 4. thieving paper; 5. base plate; 6. systemic loupus erythematosus detection zone; 7. systemic sclerosis detection zone; 8. Quality Control district.
Embodiment
Describe the present invention in detail below in conjunction with the drawings and specific embodiments.
The joint inspection test paper of embodiment 1 systemic loupus erythematosus of the present invention and systemic sclerosis
As shown in Figure 1, be the joint inspection test paper of a systemic loupus erythematosus of the present invention and systemic sclerosis.Comprise sample pad 1, the glass fibre membrane that contains colloid gold particle label 2 being closely connected with described sample pad 1 one end, the nitrocellulose filter 3 being closely connected with described glass fibre membrane 2 other ends, and the thieving paper 4 being closely connected with the other end of described cellulose membrane 3, described sample pad 1, glass fibre membrane 2, nitrocellulose filter 3 and thieving paper 4 are all arranged on base plate 5, described nitrocellulose filter 3 comprises the detection zone 6 that is coated with systemic loupus erythematosus sm recombinant antigen, be coated with the detection zone 7 and the control zone 8 that is coated with sheep anti-mouse antibody of systemic sclerosis Scl-70 recombinant antigen, described colloid gold particle label comprises micro-signal amplification system and colloid gold label mouse IgG antibody, described micro-signal amplification system is colloid gold particle-Avidin-Biotin-sm/Scl-70 recombinant antigen, wherein sm/Scl-70 recombinant antigen is systemic loupus erythematosus sm recombinant antigen or systemic sclerosis Scl-70 recombinant antigen.
A kind of human body autoantibody joint inspection test strips of the present invention, the consumption of described colloid gold particle-Avidin-Biotin-systemic loupus erythematosus sm recombinant antigen compound on glass fibre membrane 2 is 50 μ g/cm
2; The consumption of colloid gold particle-Avidin-Biotin-systemic sclerosis Scl-70 recombinant antigen compound on glass fibre membrane 2 is 55 μ g/cm
2; The consumption of colloid gold label mouse IgG antibody on glass fibre membrane 2 is 30 μ g/cm
2; The original concentration of systemic loupus erythematosus sm recombinant antigen is that the original concentration of 3.5mg/ml and systemic sclerosis Scl-70 recombinant antigen is 2.5mg/ml, concentration while being coated with the detection zone 6 of nitrocellulose membrane 3 and detection zone 7 is respectively 1.0mg/ml and 1.2mg/ml, and coated consumption is respectively 0.18 μ g/mm and 0.21 μ g/mm; Consumption when sheep anti-mouse igg antibody is coated on control zone is 0.30 μ g/mm.
The preparation method of embodiment 2 test strips of the present invention
Adopt in the present invention colloid gold particle mark.Wherein the particle diameter of colloid gold particle size is nanoscale (30～50nm), the adsorption mechanism of colloid gold particle is to utilize its electronegative character under alkali condition, with the positive charge group of protein molecule, by electrostatic attraction and in conjunction with forming immune diagnostic reagent.
Below the preparation method of test strips of the present invention is described below:
A kind of human body autoantibody joint inspection test strips, its preparation process is as follows:
1. preparation system lupus erythematosus sm recombinant antigen and systemic sclerosis Scl-70 recombinant antigen.
Consult document and with biological special software analytic system lupus erythematosus sm albumen and systemic sclerosis Scl-70 albumen, specify functional fragment and amino acid sequence thereof, obtain functional protein fragment with pcr amplification method or method for synthesizing gene, the prokaryotic expression system of employing or eukaryotic expression system are expressed target protein and are also carried out recombinant protein purification and functional evaluation according to protein characteristic.
2, prepare micro-signal amplification system
A, prepare colloid gold label Avidin: by Avidin to be marked 4 ℃ of dialysed overnight in the NaCl of 0.005mol/L pH7.0 solution in advance, to remove unnecessary salt ion, then 4 ℃ of centrifugal 1h of 100000g, remove polymkeric substance; With 0.1mol/L K
2cO
3or pH value to 9～10 of 0.1mol/L HCl adjusting collaurum liquid, mark Avidin is to form colloid gold label Avidin;
B, prepare biotinylation sm/Scl-70 recombinant antigen: be first connected with biotin and make long-armed biotin with 6-aminohexose, make again it be combined with sm/Scl-70 recombinant antigen, then under the effect of carbodiimide, itself and N-hydroxy-succinamide are contracted and, generate long-armed biotin N-maloyl imines ester (N-hydroxy-succinimido-6-biotinyl amido hexanoate, BCNHS), be biotinylation sm/Scl-70 recombinant antigen.Remove free biotin by dialysis.
C, complete micro-signal amplification system: the biotinylation sm/Scl-70 recombinant antigen that the colloid gold label Avidin that step a is obtained and step b obtain directly mixes, connect and obtain collaurum-Avidin-Biotin-sm/Scl-70 recombinant antigen compound, after washing, centrifugal treating, obtain micro-signal amplification system.
3, colloid gold label mouse IgG antibody
In the scope of pH6.5～7.0, mouse IgG and colloid gold particle protein labeling form colloid gold label mouse IgG antibody (albumen probe).
4, collaurum-Avidin-Biotin-sm/Scl-70 recombinant antigen and colloid gold label mouse IgG antibody are sprayed onto on glass fibre membrane 2, dilution parameters (dilution parameters is the technological parameter of every how many areas of ml spray solution) is 25～35cm
2/ ml, and 20～40 ℃ of temperature, humidity 10%-30%, more than 12 hours, for subsequent use by dry glass fibre membrane 2.
5, with coated damping fluid (principal ingredient is phosphate buffer, sucrose) dilution sm/Scl-70 recombinant antigen, making its concentration is 1.0mg/ml, press film liquid measure 0.18 μ l/mm, by its careful being sprayed onto uniformly on nitrocellulose filter 3 respectively, wherein nitrocellulose filter 3 apertures are 5.0～12.0 μ m, be placed in 20～40 ℃, under humidity 10%～30% condition, drying and processing is more than 12 hours, for subsequent use; With coated damping fluid dilution sheep anti-mouse igg antibody, making its concentration is 1.5mg/ml, by film liquid measure 0.20 μ l/mm, by its careful being sprayed onto uniformly on nitrocellulose filter 3, be placed in 20～40 ℃, under humidity 10%～30% condition, drying and processing is more than 12 hours, envelope, for subsequent use;
6, sample pad 1, glass fibre membrane 2, nitrocellulose filter 3 and thieving paper 4 are sequentially pasted on base plate 7, obtain test strips of the present invention.
The joint inspection test paper of systemic loupus erythematosus of the present invention and systemic sclerosis can directly use through packing, or is installed in kit, coordinates little dropper, serves as reagent card and uses.
Embodiment 3 utilizes systemic loupus erythematosus sm antibody in the ELISA test strip sample in embodiment 1 and the method for systemic sclerosis Scl-70 antibody
The present embodiment utilizes the test strips of embodiment 1 to detect the sm/Scl-70 antibody in human blood sample.
Comprise the following steps:
(1) sample of blood, drawn, the centrifugal serum of collecting.
(2) tear along aluminium foil bag cutting part, take out examination bar and keep flat, draw 5 μ l samples and vertically drip in the sample application zone (Fig. 1) shown in examination bar arrow glue; Due to capillarity, sample will be along test strips through glass fibre element film 2 and nitrocellulose filter 3.
After (3) 5 minutes, observe and show result (result showing after 30 minutes is invalid), interpretation is also recorded testing result (Fig. 2 and Fig. 3), if there is a macroscopic dark line in the detection zone 6 of nitrocellulose filter 3, show to contain in sample a large amount of systemic loupus erythematosus sm antibody, i.e. explanation is subject to proofer to suffer from systemic loupus erythematosus (systemic loupus erythematosus sm antibody positive, please also refer to Fig. 2); If there is a macroscopic dark line in detection zone 7, show to contain systemic sclerosis Scl-70 antibody in sample, illustrate and be subject to proofer to suffer from systemic sclerosis (systemic sclerosis Scl-70 antibody positive, please also refer to Fig. 2); If develop the color in detection zone 6 and detection zone 7 simultaneously, i.e. explanation is subject to proofer to suffer from systemic loupus erythematosus and systemic sclerosis simultaneously; If all there are not macroscopic colour developing lines in the detection zone 6 of nitrocellulose filter 3 and detection zone 7, show not contain in sample sm antibody or the Scl-70 antibody that can detect, illustrate and be subject to proofer not suffer from systemic loupus erythematosus and systemic sclerosis (feminine gender please also refer to Fig. 2); In the time that the detection zone 6 of sample by nitrocellulose filter 3 and detection zone 7 move to control zone 8, no matter in sample, whether contain sm antibody or Scl-70 antibody, control zone 8 all can have a dark line (C line); If control zone 8 without chromogenic line occur, illustrate test strips expired or operate wrong, testing result invalid (please refer to Fig. 3);
(4), after test finishes, test strips, application of sample pipe after using are processed by biologic medical discarded object.
Claims (9)
1. a human body autoantibody joint inspection test strips, it is characterized in that, comprise sample pad (1), the glass fibre membrane that contains colloid gold particle label (2) being closely connected with described sample pad (1) one end, the nitrocellulose filter (3) being closely connected with described glass fibre membrane (2) other end, and the thieving paper (4) being closely connected with the other end of described nitrocellulose filter (3), described sample pad (1), glass fibre membrane (2), nitrocellulose filter (3) and thieving paper (4) are all arranged on base plate (5), described nitrocellulose filter (3) comprises the detection zone (6) that is coated with systemic loupus erythematosus sm recombinant antigen, be coated with the detection zone (7) of systemic sclerosis Scl-70 recombinant antigen, and be coated with the control zone (8) of sheep anti-mouse antibody, described colloid gold particle label comprises the mouse IgG antibody of micro-signal amplification system and colloid gold label, described micro-signal amplification system is colloid gold particle-Avidin-Biotin-sm/SCL-70 recombinant antigen.
2. human body autoantibody joint inspection test strips according to claim 1, is characterized in that parallel systemic loupus erythematosus sm recombinant antigen and the systemic sclerosis SCL-70 recombinant antigen of being coated with, simultaneously detection system lupus erythematosus and systemic sclerosis.
3. human body autoantibody joint inspection test strips according to claim 2, is characterized in that, described systemic loupus erythematosus sm recombinant antigen consumption is 0.13-0.27 μ g/mm, and described systemic sclerosis SCL-70 recombinant protein consumption is 0.13-0.27 μ g/mm.
4. according to the human body autoantibody joint inspection test strips described in claim 1-3, it is characterized in that, the consumption of described colloid gold particle-Avidin-Biotin-sm/Scl-70 recombinant antigen compound is 12.5-68 μ g/cm
2.
5. human body autoantibody joint inspection test strips according to claim 1, is characterized in that, described colloid gold label has mouse IgG antibody, and consumption is 15-50 μ g/cm
2.
6. human body autoantibody joint inspection test strips according to claim 1, is characterized in that, described sheep anti-mouse antibody is sheep anti-mouse igg antibody, and the consumption of described sheep anti-mouse antibody is 0.20-0.48 μ g/mm.
7. a method of preparing human body autoantibody joint inspection test strips claimed in claim 1, is characterized in that, comprises the following steps:
(1) preparation system lupus erythematosus sm recombinant antigen, systemic sclerosis Scl-70 recombinant antigen and colloid gold label mouse IgG antibody;
(2) prepare micro-signal amplification system
A, prepare colloid gold label Avidin
By Avidin to be marked dialysed overnight in the NaCl of 0.001～0.01mol/L solution in advance, centrifugal, pH value to 9～10 of adjusting collaurum liquid, mark Avidin is to form colloid gold label Avidin;
B, prepare biotinylation sm recombinant antigen and biotinylation Scl-70 recombinant antigen according to a conventional method;
C, complete micro-signal amplification system
The biotinylation sm/Scl-70 recombinant antigen that the colloid gold label Avidin that step a is obtained and step b obtain mixes, and connects and obtains collaurum-Avidin-Biotin-sm/Scl-70 recombinant antigen compound, and washing, obtains micro-signal amplification system;
(3) by dilution parameters 25～40cm
2/ ml, is sprayed onto the collaurum-Avidin-Biotin of step (2)-sm recombinant protein and collaurum-Avidin-Biotin-Scl-70 recombinant protein on glass fibre membrane, dry more than 12 hours, for subsequent use;
(4) with coated damping fluid dilution sm recombinant antigen, making its concentration is 1.0～1.5mg/ml, by film liquid measure 0.15～0.2 μ l/mm, by its careful being sprayed onto uniformly on nitrocellulose filter, dry more than 12 hours, for subsequent use;
With coated damping fluid dilution Scl-70 recombinant antigen, making its concentration is 1.0～1.5mg/ml, by film liquid measure 0.15～0.20 μ l/mm, by its careful being sprayed onto uniformly on nitrocellulose filter, dry more than 12 hours, for subsequent use;
With coated damping fluid dilution sheep anti-mouse antibody, making its concentration is 1.0～2.0mg/ml, by film liquid measure 0.16～0.20 μ l/mm, by its careful being sprayed onto uniformly on nitrocellulose filter, dry more than 12 hours, for subsequent use;
(5) glass fibre membrane of sample pad, step (3), the nitrocellulose filter of step (4), thieving paper are pasted on base plate according to the order of sequence, obtain final product.
8. the preparation method of human body autoantibody joint inspection test strips according to claim 7, is characterized in that, in described step (3), the consumption of collaurum-Avidin-Biotin-sm recombinant antigen compound is 12.5-68 μ g/cm
2, the consumption of collaurum-Avidin-Biotin-Scl-70 recombinant antigen compound is 12.5-68 μ g/cm
2.
9. the preparation method of human body autoantibody joint inspection test strips according to claim 7, is characterized in that, in described step (4), sm recombinant antigen consumption is 0.13-0.27 μ g/mm; The consumption of Scl-70 recombinant antigen is 0.13-0.27 μ g/mm; The consumption of described sheep anti-mouse antibody is 0.20-0.48 μ g/mm.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310007578.5A CN103913568A (en) | 2013-01-04 | 2013-01-04 | Joint inspection test paper strip for human body autoantibody and preparation method thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109900909A (en) * | 2019-02-28 | 2019-06-18 | 中国科学院广州生物医药与健康研究院 | A nano-gold-labeled lateral flow immunochromatographic test strip for the detection of osteopontin |
| CN110261624A (en) * | 2019-07-30 | 2019-09-20 | 江苏省人民医院（南京医科大学第一附属医院） | Fluorescence immunochromatography reagent strip for determining human blood phosphotyrosine adaptor protein ShcA antibody and preparation method and application thereof |
| CN111868528A (en) * | 2018-04-03 | 2020-10-30 | 赛诺菲 | Lateral flow immunoassay strip device |
-
2013
- 2013-01-04 CN CN201310007578.5A patent/CN103913568A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111868528A (en) * | 2018-04-03 | 2020-10-30 | 赛诺菲 | Lateral flow immunoassay strip device |
| CN109900909A (en) * | 2019-02-28 | 2019-06-18 | 中国科学院广州生物医药与健康研究院 | A nano-gold-labeled lateral flow immunochromatographic test strip for the detection of osteopontin |
| CN110261624A (en) * | 2019-07-30 | 2019-09-20 | 江苏省人民医院（南京医科大学第一附属医院） | Fluorescence immunochromatography reagent strip for determining human blood phosphotyrosine adaptor protein ShcA antibody and preparation method and application thereof |
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Application publication date: 20140709 |