CN1147729C - Semi-quantitative one-step immunologic diagnosis method - Google Patents
Semi-quantitative one-step immunologic diagnosis methodInfo
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- CN1147729C CN1147729C CNB981228925A CN98122892A CN1147729C CN 1147729 C CN1147729 C CN 1147729C CN B981228925 A CNB981228925 A CN B981228925A CN 98122892 A CN98122892 A CN 98122892A CN 1147729 C CN1147729 C CN 1147729C
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Abstract
The present invention relates to a semiquantitative one-step immunologic diagnosis method. Colors are displayed on specific carriers of antibodies containing colloidal gold, etc. by a colloidal gold technique and the reaction principle of required test components (namely antigens) and antibody phases, and comparison is carried out according to the depth of a color band and the color depth of a color band of a single cloning specificity granular body A marked by a solidification gold granule to determine the concentration range of the antigens. The semiquantitative one-step immunologic diagnosis method is very simple and convenient. Only immune needs to be inserted for 3 to 5 minutes, and a result can be obtained. The result is satisfied by a clinic test.
Description
Technical field
The present invention relates to a kind of sxemiquantitative gold-marking immunity chromatography test-strips, the test-strips of more particularly utilizing antigen, antibody phase reaction principle on specific carrier, to detect with the colour developing depth.
Background technology
Dot immunobinding assay is that solid phase labelling immunoassays new technology, its principle that the mid-80 grows up is to be carrier with the miillpore filter fast, by diafiltration or capillarity antigen-antibody reaction is carried out fast, finish in the testing process number minute, painted spot appears in positive reaction on film.What set up at first is the immunity percolation test of enzyme labeling, thereafter with coloured particulates such as collaurum thing that serves as a mark, not only reduced operation steps, and reagent can be in the room temperature long preservation, as: be the easy detection enzyme immunoassay (EIA) commonly used that carries out the antibody (comprising anti-HBcIgG and anti-HBcIgM) of resistance of hepatitis B (hepatitis B) virus core antigen (HBcAg) apace.After this scientist's detection method of resisting HBc is improved and has been set up the simple and rapid golden immune HBcAg of dripping (dripping golden method).What used at that time is the indirect method pattern, and envelope antigen is expensive high-purity saccharomycete reorganization HBcAg.Homemade HBcAg is impure more, occurs false positive when on probation.After again after further study, set up the golden method of dripping of the anti-HBc IgM of detection of prize law pattern and IgG antibody, through Preliminary Applications, effect is satisfied.
The immunity-chromatography test of new development is fixed on various reaction reagent branches on the same strip, detects the end that sample is added in strip, and with a kind of agent dissolves, dividing a word with a hyphen at the end of a line by capillarity contacts with another kind of reagent on the film, reacts.Become a reagent, one with regard to simplification like this and go on foot the quick diagnosis test of finishing.The simplest reagent is just as measuring the test strips of glucose in urine in the biochemical investigation, the leaching sample gets final product observations, and the characteristics of this test are single part of mensuration, and are easy, quick, except that commercially available reagent, do not need any instrument and equipment, therefore be specially adapted to small-sized laboratory, doctor clinic and emergency treatment check.The early pregnancy diagnosis reagent of most widely used first-elected detection urine human chorionic gonadotrophin (HCG) is easy and simple to handle in clinical examination, self testing at home.Secondly be infectious disease antigen or detection of antibodies, existing multiple commercially available reagent supply.
The immunohistochemistry new technology that though immune technology for gold is a development in recent years to get up.Along with developing rapidly of immunocytochemical technique, optimize more responsive, safety and reliable new method in order constantly to explore, scholars are seeking better label always.In technical developments such as immunofluorescence, immuno-enzymatic, radio-immunity, immune colloidal gold technique more and more is subjected to vast immunocytochemistry person's attention.But above-mentioned the whole bag of tricks remains in certain defective, and can adopt single stage method to carry out immune detection at present mainly is the qualitative sxemiquantitative or quantitative of still can not accomplishing.
Summary of the invention
The objective of the invention is to seek the fast and convenient gold-marking immunity chromatography test-strips that is had half-quantitative detection antigen concentration scope " chemical examination composition " (promptly the claiming antigen) in blood, urine, the saliva of a kind of energy.
The present invention adopts colloidal gold technique need to make it the principle of " chemical examination composition " (being antigen) and antibody phase reaction, on the specific support that contains antibody such as collaurum, demonstrate color and do one relatively, decision antigen concentration scope according to the colour band depth and the shade that solidifies goldc grains mark monoclonal specific antibody A colour band.Adopt colloidal gold technique be with collaurum as a kind of label, it can be stablized and adhesion protein and the biologically active of albumen does not have obvious change promptly.Collaurum is meant the aqueous solution of gold, and it has the characteristic of common sol.The color of colloidal sol depends on material dispersed color.Concerning with a kind of aqueous solution of material, particle size difference, the color that presents are also different, and generally being applied to immunohistochemical colloid gold particle is to present redness in the 5-60nm scope.(α-FP) antibody, liver surface antigen (BHsAg), myocardium calcium protein (Troponine) combine collaurum with enough human chorionic gonadtropin (hCG) antibody, lutropin (LH) antibody, embryo cancer antigen (CEA) antibody, alpha-fetoprotein, it keeps red constant, this collaurum that is labelled protein, again this collaurum bag is drained by vacuum on all-glass paper, made the 3rd district immobilization goldc grains mark monoclonal specific antibody A.On nitrocellulose filter, rule respectively at the rabbit anti-mouse igg that is coated with purifying on the nitrocellulose filter or sheep anti-mouse igg and anti-α-hCG antibody, anti-FSH antibody, anti-LH antibody, anti-CEA antibody, α-FP antibody, TSH antibody, PSA antibody, HBsAg antibody and myocardium calcium antibody etc., bag was cut into little in 20 minutes by placement in rearmounted 37 ℃ of baking ovens makes the 4th district immobilization monoclonal specific antibody B (T district) respectively, the 5th district's immobilization sheep anti-mouse antibody or rabbit anti-mouse antibody (C district).
Second district adopts blocking agent to combine a neutralization reaction by the antigen in the specimen with antibody in the blocking agent, and " blocking agent " can be antibody, protein, amino acid or other chemical substances.Available from Canadian Gellix company, product is called Geni Block reagent.
First district is a glass layer, and the 6th district is a thieving paper.
The preparation (see figure 2) of test strips or box: get the plastic plate that a 10mm PVC simultaneously posts double faced adhesive tape, glass fibre is sticked on the bottom of plastic plate, as first district.
The blocking agent of second district for buying from the market.
The 3rd district is colloid gold label β-hCG antibody or FSH antibody or LH antibody or CEA antibody or α-FP antibody or liver surface antigen or myocardium calcium protein antibody, can change the different protein antibodies of colloid gold label as the 3rd district according to the requirement of test test agent.
The 4th district is that bag is by anti-α-hCG, anti-FSH, anti-LH, anti-CEA, anti-α-FP, anti-TSH, anti-HbsAg, anti-myocardium protein phase antagonist on the nitrocellulose membrane, and used different anti-protein antibodies mainly is to require to do corresponding variation with the 3rd district according to specimen.
Wrap by rabbit anti-mouse igg or sheep anti-mouse igg on the nitrocellulose membrane in the 5th district.
The 6th district is the suction scraps of paper, and unnecessary moisture content is blotted.
Test substance can be an antigen, also can be antibody as hCG, LH, FSH, CEA, α-FP, TSH, PSA, HBsAg, myocardium calcium protein etc., also can be other compounds.
Testing sample drips at " paper slip " first district-glass fibre, and liquid is absorbed immediately, and is up with capillary theory, liquid dissolving to be measured second district " blocking-up " reagent, and its antigen that comprises combines-neutralization reaction with " blocking-up " reagent." blocking-up " amount of reagent is high more, in and the conjugated antigen amount many more.In conjunction with the ability that combines again of antigen forfeiture and antibody A and B.Liquid goes upward to the 3rd district, dissolve gold labeling antibody A, not in and antigen and golden labeling antibody A formation immune complex-purple rugose rose (Rosa rugosa var look.This purple liquid continues up, during to the T district, forms " antibody B-Ag-Ab A-goldc grains " precipitation line.Precipitation line shade and antigen amount, " blocking-up amount of reagent " have linear relationship.When purple liquid continues to go upward to the 5th district-" C district ", meet with " C district " immobilization sheep anti-mouse antibody or rabbit anti-mouse antibody, because of monoclonal antibody all belongs to cloned mouse antibody, so no matter whether antigen exists, " C district " is always have precipitation line appearance-" a sheep anti-mouse antibody mouse monoclonal antibody goldc grains ".This colour band is generally as quality control: purple band do not occur as " C district ", " paper tape " inactivation then is described, test must be reformed.Among the present invention, " C district " is except that doing quality control, also as " sxemiquantitative contrast band ", the i.e. certain antigen concentration content of the color depth in " C district " representative.
In the time of testing setup, during as 5 minutes or 10 minutes, with the colour band depth in " T district " and " C district " color relatively, to determine the antigen concentration scope.
Description of drawings:
Fig. 1 be on the nitrocellulose membrane bag by rabbit anti-mouse antibody or sheep anti-mouse antibody and anti-α-hCG antibody or anti-FSH antibody or anti-LH antibody or anti-CEA antibody or anti-α-FP antibody or PSA antibody or anti-HBsAg antibody or anti-myocardium calcium protein antibody wherein 1 for bag by district's band of district's band of rabbit anti-mouse antibody or sheep anti-mouse antibody.
2 for wrapping by district's band of anti-α-hCG antibody.
Fig. 2 is a semi-quantitative one-step method immunodiagnosis test paper
3 is first district: the absorption of sample district
4 is second district: immobilization blocking-up reagent
5 is the 3rd district: immobilization goldc grains mark monoclonal specific antibody A
6 is the 4th district: immobilization monoclonal specific antibody B (T district)
7 is the 5th district: immobilization sheep anti-mouse antibody or rabbit anti-mouse antibody C district
8 is the 6th district: the suction scraps of paper
Embodiment
The making of embodiment 1 application testing bar semiquantitative determination human chorionic gonadtropin hCG content measuring bar in urine:
One material
1. rabbit anti-mouse igg (affinity purification): provide by Shanghai branch office of magnificent biotech firm
2.hCG standard items: produce by the biochemical-pharmaceutical factory, Shanghai
3. the suction scraps of paper: that adopts that Shanghai papermaking research institute produces copies the quantitative thieving paper of sheet
4. anti-β-hCG antibody, anti-α-hCG antibody, Buffer F, collaurum, nitrocellulose filter, all-glass paper etc. provide by the U.S..
5. freshly voided urine sample: provide by Rui Jin hospital of Shanghai Second Emdical University
6. " hCG blocking agent " (buying) by Vancouver, CAN Gellix company
Two methods:
1. colloid gold label β-hCG antibody: get the β-hCG antibody of required collaurum and respective amount respectively,, make its combination, add 10% BSA used as stabilizers stirring under the vibration.Remove free unconjugated β-hCG antibody and fully unstable gold grain and its condensation product with centrifuge method.The peony precipitation is collaurum-anti-β-hCG antibody complex in the centrifuge tube bottom.
2. purifying:
Use the polyglycol washing precipitate, the centrifugal supernatant that goes gets the peony precipitation.
3. the precipitation part is dissolved loose red precipitate with buffer.The shop all-glass paper, vacuum is drained.
4. bag is cut into nitrocellulose filter the size of 0.5 * 10cm, uses the rabbit anti-mouse igg and the anti-α-hCG of purifying to rule (as shown in the figure) on nitrocellulose filter, the rearmounted 37 ℃ of baking ovens of bag quilt 20 minutes.
5. get one and be of a size of the plastic bottom board that the thick about 0.5mmd of 10cm * 12cm simultaneously posts double faced adhesive tape, respectively filter paper, the golden millimeter paper that is marked with β-hCG antibody, the nitrocellulose filter that is coated with the anti-mouse of rabbit and anti-α-hCG antibody and filter paper are made by method shown in Figure 2.
300,1200,20000mI μ/ml is divided into three groups on A, B, C type with in first districts insertion urine of above-mentioned test-strips (urine must not surpass the 3rd district) by its blocking-up dosage 1,5.0,25.0 μ l differences with test-strips set three critical values according to the hCG content in the urine samples:, after treating 5 minutes, urine upwards slowly moves under the chromatography effect, upwards penetrates on the nitrocellulose filter being marked at the anti-β-hCG antibody dissolving on the all-glass paper and being dynamic and driving it.If urine is positive, promptly contain in the urine hCG hormone and 〉=during 25mIU/ml, because the bridging effect of anti-β-hCG antibody-hCG-α-hCG antibody-gold is coated with α-hCG antibody part and promptly shows the distinctive redness of collaurum on the film.Urine-colorant continues infiltration up, arrives the position that is coated with the rabbit anti-mouse antibody.This district's band promptly shows red, is control line.When hCG in the urine 〉=300mIU/ml, A type strip is to present two red zones on the nitrocellulose filter, is positive, as a red zone, i.e. hCG≤300mIU/ml in the urine only occurring.
Experimental result is calculated (test duration: 5 minutes) as shown in table 1
Table 1:
| The paper slip model | Critical value mIU/ml | " blocking-up reagent " dosage μ l | C district typical value mIU/ml | Experimental result mIU/ml |
| A | 300 | 1.0 | 300 | As T＞C, concentration＞300 then |
| B | 1,200 | 5.0 | 1,200 | As T＞C, concentration＞1,200 then |
| C | 20,000 | 25.0 | 20,000 | As T＞C, concentration＞20,000 then |
Test one:
3050mIU/ml is diluted to negative urine with the hCG standard items, is mixed with 300,600,1,200 and 20, and 000mIU/ml with this negative urine, detects with paper slip A type, Type B, C type, and the result classifies table 2 as.
HCG standard items 3,050mIU/ml, with normal urine be diluted to 150,300,600,12K, 2.4K, 10K, 20K, 40K mlU/ml solution.
Table 2:
Test findings (5 minutes)
| The model critical value | A 300mIU/ml | B 1.2 mIU/ml | C 20KmIU/ml |
| HCG concentration | A type hCG concentration | Type B hCG concentration | The C type |
| 150mIU/ml 300mIU/ml 600mIU/ml | T＜C 600mIU/ml T＝C 1.2KmIU/ml T＞C 2.4KmIU/ml | T＜C 10KmIU/ml T＝C 20KmIU/ml T＞C 40KmIU/ml | T＜C T＝C T＞C |
Test two:
From clinically, divide and collect pregnant woman's urine sample 29,30 and 35 examples for three times.Compare detection with enzyme mark method and single stage method sxemiquantitative paper slip, the result is as shown in table 3.Enzyme linked immunosorbent assay and sxemiquantitative paper strip method correlation ratio are respectively for three times:
Table 3-1: clinical testing 1
| Object number .. to be measured | Enzyme-linked method mIU/ml | A type 300 critical values | Type B 1.2K critical value | C type 20K critical value |
| 001 | 51K | T＞＞C | T＞＞C | T＞＞C |
| 008 | 135 | T＝/＜C | T＜C | - |
| 010 | 24K | T＞＞C | T＞＞C | T＞C |
| 011 | 2,050 | T＞C | T＝/＞C | - |
| 012 | 1,200 | T＞C | T＝C | - |
| 020 | 410 | T＝C | T＜C | - |
| 023 * | 70K | T＝C | T＝C | T＝C |
| 048 | 60K | T＞＞C | T＞＞C | T＞＞C |
| 057 | 2,000 | T＞＞C | T＝C | - |
| 061 | 8 | - | - | - |
| 062 | 500 | T＞C | T＜C | - |
| 079 | 2,100 | T＞＞C | T＞C | T＜C |
| 083 | 2,300 | T＞＞C | T＞C | T＜＜C |
| 084 | 3,900 | T＞＞C | T＞C | T＜C |
| 100 | 2,350 | T＞＞C | T＞C | T＜C |
| 101 | 10 | - | - | - |
| 102 | 3,300 | T＞＞C | T＞C | T＜C |
| 123 | 13,000 | T＞＞C | T＞＞C | T＜C |
| 147 | 825 | T＞＞C | T＞C | T＜C |
| 182 | 155 | T＜C | - | - |
| 194 | 265 | T＝C | T＜＜C | - |
| 199 | 14K | T＞＞C | T＞＞C | T＞C |
| 201 | 1,550 | T＞C | T＞C | T＜＜C |
| 214 | 17K | T＞＞C | T＞＞C | T＝/＞C |
| 236 | 2,900 | T＞＞C | T＞＞C | T＜C |
| 240 | 1,000 | T＞＞C | T＝C | - |
| 243 | 5,750 | T＞＞C | T＞＞C | T＜C |
| 247 | 5,250 | T＞＞C | T＞＞C | T＝C |
| 251 | 12 | T＜＜C | - | - |
| Add up to | Correlativity | |||
| 29 | 100％ | 96.5％ | 93.1％ | |
Attention: the no p-wire of "-" expression, No. 23 case is used
ClomitTreatment.
Table 3-2:
Clinical testing 2
| Object No. to be measured | Enzyme-linked method mIU/ml | A type 300 critical values | Type B 1.2K critical value | C type 20K critical value |
| 04 | 28K | T＞C | T＞C | T＞C |
| 05 | 21.5K | T＞C | T＞C | T＞C |
| 06 | 474 | T＞C | T＜C | - |
| 07 | 3.3K | T＞C | T＞C | - |
| 08 | 1.2K | T＞C | T＝C | - |
| 09 | 6.4K | T＞C | T＞C | T＜＜C |
| 10 | 5.4K | T＞C | T＞C | T＜＜C |
| 11 | 9K | T＞C | T＞/＝C | - |
| 12 | 63K | T＞C | T＞C | T＞C |
| 13 | 2.4K | T＞C | T＞/＝C | - |
| 23 | 28 | - | - | - |
| 24 | 25K | T＞C | T＞C | T＞C |
| 25 | 2.7K | T＞C | T＞C | T＜C |
| 26 | 24K | T＞C | T＞C | T＞/＝C |
| 27 | 3K | T＞C | T＞C | - |
| 28 | 4K | T＞C | T＞/＝C | - |
| 29 | 2.9K | T＞C | T＞C | T＜＜C |
| 30 | 4.0K | T＞C | T＞C | T＜＜C |
| 31 | 1.8K | T＞C | T＞C | - |
| 32 | 21.6K | T＞C | T＞C | T＞C |
| 33 | 23K | T＞C | T＞C | T＞C |
| 34 | 2.6K | T＞C | T＞C | T＜＜C |
| 36 | 19K | T＞C | T＞C | T＞C |
| 37 | 12K | T＞C | T＞C | T＝C |
| 38 | 20K | T＞C | T＞C | T＝C |
| 39 | 11.8K | I＞C | T＞C | T＜C |
| 41 | 7.6K | T＞C | T＞C | T＜C |
| 47 | 30 | T＜C | - | - |
| 48 | 3.2K | T＞C | T＞C | T＜C |
| 50 | 25.5K | T＞C | T＞C | T＜C |
| Add up to | Correlativity | |||
| 100％ | 90.0-100％ | 96.6％ | ||
Table 3-3
Clinical testing 3
Lot number 9804
| Object No. to be measured | Enzyme-linked method mIU/ml | A type 300 critical values | Type B 1.2K critical value | C type 20K critical value |
| 01 | 3.1K | T＞C | T＞C | T＜C |
| 02 | 3.2K | T＞C | T＞C | - |
| 03 | 5.6K | T＞C | T＞C | T＞C |
| 04 | 1.1K | T＞C | T＝C | - |
| 05 | 48.0K | T＞C | T＞C | T＞C |
| 06 | 3.6K | T＞C | T＞C | T＜C |
| 07 | 2.8K | T＞C | T＞C | T＜C |
| 08 | 23.5K | T＞C | T＞C | T＞/＝C |
| 09 | 9.4K | T＞C | T＞C | T＞C |
| 10 | 2.4K | T＞C | T＞C | T＜C |
| 11 | 6.2K | T＞C | T＞C | T＞C |
| 12 | 61.0K | T＞C | T＞C | T＞C |
| 13 | 8.0K | T＞C | T＞C | T＜C |
| 14 | 38.0K | T＞C | T＞C | T＞C |
| 15 | 6.0K | T＞C | T＞C | - |
| 17 | 8.6K | T＞C | T＞C | T＝C |
| 18 | 1.1K | T＞C | T＝C | T＜C |
| 19 | 1.1K | T＞C | T＝C | T＜C |
| 20 | 4.3K | T＞C | T＞C | - |
| 21 | 4.4K | T＞C | T＞C | T＜C |
| 22 | 2.7K | T＞C | T＞C | T＜C |
| 23 | 28.4K | T＞C | T＞C | T＞C |
| 24 | 6.6K | T＞C | T＞C | T＜C |
| 25 | 24.0K | T＞C | T＞C | T＞C |
| 26 | 32.0K | T＞C | T＞C | T＞C |
| 27 | 5.0K | T＞C | T＞C | T＜C |
| 28 | 12.0K | T＞C | T＞C | T＜C |
| 29 | 5.4K | T＞C | T＞C | T＜C |
| 30 | 1.3K | T＞C | T＝C | - |
| 31 | 2.8K | T＞C | T＞C | - |
| 32 | 20.8K | T＞C | T＞C | T＞C |
| 33 | 3.0K | T＞C | T＞C | - |
| 34 | 6.1K | T＞C | T＞C | T＜C |
| 35 | 5.6K | T＞C | T＞C | - |
| 36 | 5.5K | T＞C | T＞C | - |
| Add up to | Correlativity | |||
| 35 | 100％ | 100％ | 91.4％ | |
Enzyme linked immunosorbent assay and sxemiquantitative paper strip method correlation ratio are respectively table 3 three times
Table 3
| The sxemiquantitative paper strip method | ||||
| The A type | Type B | The C type | ||
| Enzyme | 29 examples for the first time | 100％ | 96.5％ | 93.1％ |
| Mark | 30 examples for the first time | 100％ | 90～100％ | 96.6％ |
| Method | 35 examples for the first time | 100％ | 100％ | 91.4％ |
Test three:
From clinically, divide secondary to collect each 25 example of pregnant woman's urine sample.Compare detection with sheep red blood cell method and single stage method sxemiquantitative paper slip, the result is as shown in table 4.
Table 4
Clinical trial 4
Lot number 9801
| Object No. to be measured | Enzyme-linked method mIU/ml | A type 300 critical values | Type B 1.2K critical value | C type 20K critical value |
| 01 | ＜312 | T＜C | T＜C | T＜C |
| 02 | ＜312 | T＜C | T＜C | T＜C |
| 03 | ＞312 | T＞C | T＞C | T＜C |
| 04 | ＞312 | T＞C | T＜C | T＜C |
| 05 | ＞625 | T＞C | T＜C | T＜C |
| 06 | ＞625 | T＞C | T＜C | T＜C |
| 07 | 1250 | T＞C | T＞C | T＜C |
| 08 | 5000 | T＞C | T＞C | T＜C |
| 09 | 10000 | T＞C | T＜C | T＜/＝C |
| 10 | 20K | T＞C | T＞C | T＞C |
| 11 | 40K | T＞C | T＞C | T＞C |
| 12 | 80K | T＞C | T＞C | T＞C |
| 13 | ＞160K | T＞C | T＞C | T＞C |
| 14 | ＜312 | T＜C | T＜C | T＜C |
| 15 | 312 | T＞C | T＜C | T＜C |
| 16 | 625 | T＜C | T＜C | T＜C |
| 17 | 1250 | T＞C | T＞C | T＜C |
| 18 | 2500 | T＞C | T＞C | T＜C |
| 19 | 5000 | T＞C | T＞C | T＜C |
| 20 | 10K | T＞C | T＞C | T＜C |
| 21 | 20K | T＞C | T＞C | T＞C |
| 22 | 20K | T＞C | T＞C | T＞C |
| 23 | 10K | T＞C | T＞C | T＜C |
| 24 | 160K | T＞C | T＞C | T＞C |
| 25 | 160K | T＞C | T＞C | T＞C |
| Correlativity | ||||
| Add up to 25 | 96％ | 100％ | 100％ | |
Sheep red blood cell method and sxemiquantitative paper strip method correlation ratio are:
| The sxemiquantitative paper strip method | ||||
| The A type | Type B | The C type | ||
| Sheep red blood cell method | 25 examples | 96％ | 100％ | 100％ |
Embodiment 2. application testing bar sxemiquantitative lutropin LH content in urine
This test-strips is to detect the concentration range of lutropin LH in the urine, and material therefor is with hCG sxemiquantitative test-strips, just used pair of L H monoclonal antibody (self-control) and " LH blocks reagent " (GelIix, Vancouva, Canada).The critical value of paper slip model A, B, C is respectively 300,500,1, and 000mIU/ml is diluted in the negative urine of LH with LH mark liquid, is mixed with the positive urine sample to be measured of variable concentrations LH, and test result is as shown in table 5.
Table 5:
| The test-strips model | Critical value mIu/ml | LH blocking-up reagent dosage μ l | C district typical value mIu/ml | Solution preparation value mIu/ml to be measured | Test result |
| A | 300 | 3.0 | 300 | 1500 400 | T＜C T＞C |
| B | 500 | 6.0 | 500 | 400 600 | T＜C T＞C |
| C | 1,000 | 12.0 | 1,000 | 800 1,200 | T＜C T＞C |
Embodiment 3
This testing cassete of prostate-specific-antigen PSA sxemiquantitative testing cassete is a concentration range of surveying prostate specific antigen Prostate Specific Ait:gen (PSA) in the blood.Material therefor be a pair of PSA monoclonal antibody (self-control) and blocking-up reagent Geni Blockreagent (Gellix, Vancouva, Canada).Testing cassete is A and Type B.
Table 6
| The testing cassete model | Critical value ng/ml | Blocking-up reagent dosage μ l | C district typical value ng/ml | Solution ng/ml to be measured | Test result |
| A | 4 | 0 | 4 | 2 4 6 | T＜C T＝C T＞C |
| B | 20 | 0.5 | 20 | 15 20 25 | T＜C T＝C T＞C |
Embodiment 4
This testing cassete of thyroid-stimulating hormone TSH sxemiquantitative testing cassete is the concentration range that detects thyroid-stimulating hormone TSH in the blood.Whether the TSH level is too high to determine object to be measured-adult or baby.Material therefor, test site and golden labeling antibody are a pair of TSH monoclonal antibody (self-control), check plot C
1And C
2Be the sheep anti mouse polyclonal antibody.C
1Represent low dosage " sxemiquantitative contrast band ", C
2Represent high dose " sxemiquantitative contrast band ".
Table 7
| The testing cassete critical value setting | Solution to be measured | Test result |
| C 1 20μIU/ml C 2 60μIU/ml | 15 20 25 55 60 65 | T＜C 1 C 1＝T＜C 2 C 1＜T＜C 2 C 1＜T＜C 2 C 1＜T＝C 2 C 2＜T |
Claims (5)
1, a kind of sxemiquantitative gold-marking immunity chromatography test-strips, from bottom to top comprise absorption of sample district, first district, the 3rd district immobilization goldc grains mark specific antibody A, the corresponding specific antibody B of the 4th district's immobilization and A is the T district, the 5th district's immobilization non-specific antibody is the C district, thieving paper district, the 6th district, it is characterized in that between first district and the 3rd district, also comprising second district immobilization blocking-up reagent area, immobilization blocking-up reagent can be with the testing sample combination, described C district is quality control and sxemiquantitative contrast band, the antigen critical value concentration that its reacted shade representative is set in advance, the color in T district, reaction back and the concentration of antigen are linear, after reaction, the shade in T district is compared with C district color, but the antigen concentration scope is measured in sxemiquantitative.
2, sxemiquantitative gold-marking immunity chromatography test-strips according to claim 1 is characterized in that immobilization blocking-up reagent is antibody, protein, amino acid.
3, sxemiquantitative gold-marking immunity chromatography test-strips according to claim 1 is characterized in that immobilization blocking-up reagent area can also incorporate the 3rd district into.
4, sxemiquantitative gold-marking immunity chromatography test-strips according to claim 1 is characterized in that test substance can be an antigen, comprises hCG, LH, FSH, CEA, α-FP, TSH, HbsAg, PSA, myocardium calcium protein, also can be antibody.
5, a kind of sxemiquantitative gold-marking immunity chromatography diagnosis box, it is characterized in that comprising a kind of sxemiquantitative gold-marking immunity chromatography test-strips, this test-strips from bottom to top comprises absorption of sample district, first district, second district immobilization blocking-up reagent area, the 3rd district immobilization goldc grains mark specific antibody A, the corresponding specific antibody B of the 4th district's immobilization and A is the T district, the 5th district's immobilization non-specific antibody is the C district, thieving paper district, the 6th district, described C district is quality control and sxemiquantitative contrast band, the antigen critical value concentration that its reacted shade representative is set in advance, the color in T district, reaction back and the concentration of antigen are linear, after reaction, the shade in T district is compared with C district color, but the antigen concentration scope is measured in sxemiquantitative.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB981228925A CN1147729C (en) | 1998-12-30 | 1998-12-30 | Semi-quantitative one-step immunologic diagnosis method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB981228925A CN1147729C (en) | 1998-12-30 | 1998-12-30 | Semi-quantitative one-step immunologic diagnosis method |
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| Publication Number | Publication Date |
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| CN1260490A CN1260490A (en) | 2000-07-19 |
| CN1147729C true CN1147729C (en) | 2004-04-28 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB981228925A Expired - Fee Related CN1147729C (en) | 1998-12-30 | 1998-12-30 | Semi-quantitative one-step immunologic diagnosis method |
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| CN (1) | CN1147729C (en) |
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| CN1892223B (en) * | 2005-04-11 | 2012-08-22 | 兰州大学 | Colloidal gold semi-quantitative quick immunity diagnosis test-paper stripe |
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- 1998-12-30 CN CNB981228925A patent/CN1147729C/en not_active Expired - Fee Related
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1896740B (en) * | 2005-07-15 | 2012-05-23 | 兰州大学 | Colloidal golden test-paper strip for fastly testing oxidative low-density fatty protein by semi-quantitative method and its production thereof |
| CN102183663A (en) * | 2011-03-24 | 2011-09-14 | 武汉璟泓万方堂医药科技有限公司 | Qualitative semi-quantitative dual-purpose female ovulation hormone detection test paper and colorimetric card |
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|---|---|
| CN1260490A (en) | 2000-07-19 |
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Address after: San Diego Applicant after: Lu Laboratory Corp. Inventor after: Lu Weizhao Applicant after: Shanghai Institute of Materia Medica, Chinese Academy of Sciences Inventor after: Lu Jingyi Inventor after: Gu Zhiping Inventor after: Cao Lin Address before: No. 294, Taiyuan Road, Shanghai Applicant before: Shanghai Institute of Materia Medica, Chinese Academy of Sciences Inventor before: Gu Zhiping Inventor before: Lu Weizhao Inventor before: Cao Lin |
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