CN102628867A - Double antibody latex enhanced retinol binding protein detection kit - Google Patents
Double antibody latex enhanced retinol binding protein detection kit Download PDFInfo
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Abstract
The invention relates to a double antibody latex enhanced retinol binding protein detection kit. More specifically, the invention discloses a double antibody coating latex enhanced immunoturbidimetry kit for detecting retinol binding protein. The kit contains a reagent 1, a reagent 2 and a calibrator. Paring monoclonal antibody A and B are respectively coated on latex particles. Coated antibody is bonded with RBP to be detected, and a plurality of bonders are aggregated together to form detectable turbidity change. The detection kit provided by the invention has high sensitivity, can be used to detect urine samples and serum samples and can be used as nutritive index to detect RBP in serum. Simultaneously, through the detection of RBP in urine, the kit is sensitive to the damage degree of renal proximal tubule.
Description
Technical field
The present invention relates to a kind of double antibody latex and strengthen the Retinal-binding protein detection kit; More specifically; The invention discloses a kind of two anti-turbid kit of latex enhancing immune transmittance that encapsulates that detects RBP ELISA, it comprises reagent 1 and reagent 2 and calibration object.
Background technology
(retinol binding protein RBP) is made up of a polypeptied chain and fraction carbohydrates RBP ELISA, does not contain neutral sugar and amidohexose, is made up of 182 amino acid; Molecular weight is 21KD; Isoelectric pH 4.4-4.8, half life period 3-12h.
RBP family is divided into 5 types, comprises the interior RBP of RBP, the nuclear receptor of retinoic acid, the specific extracellular RBP of visual organization and the specific cell of visual organization in secreted RBP, the cell.
RBP is distributed widely in normal person's body fluid, has the function of transhipment retinol to surrounding tissue from liver cell.RBP mainly exists with the composite form that retinol, prealbumin combine in the blood, when the retinol in the compound with after target cell combines, RBP just separates with prealbumin, leaches from glomerulus, by the absorption of near-end renal cells, degrade.So when obstacle appearred in the heavy absorption function of renal tubule, RBP content raise in the urine.There is article to show that RBP is the sensitive indicator of reflection kidney proximal tubule damage in the urine through comparing with indexs such as microdose urine protein, urea, creatinines, can occurs unusual by first sundry item.The range of normal value of RBP is to be no more than 0.7mg/L in the urine, requires detection kit to have good sensitivity.
RBP is mainly synthetic in liver cell; Carrier protein as retinol; In liver, combine at 1: 1 with retinol; 90% RBP combines formation holo-RBP with retinol in the body, combines the formation three-dimensional composite with prealbumin with 1: 1 after being released into blood, transports retinol to surrounding tissue target cell jointly.Have only holo-RBP just can go into blood, when protein malnutrition, the protein synthesis of liver is not enough, and causes that RBP is not enough, and retinol can not get into blood, and retinol plays regulating action to the secretion of RBP, both being proportionate property.Participate in the transhipment of vitamin A with prealbumin, serum RBP and serum vitamin A content have high correlation, think that RBP can be used as the sensitive indicator of reflection VAD.Analysis nutritive disease patient's plasma proteins changes the variation of discovery blood plasma RBP early than albumin and transferrins; And be higher than albumin and transferrins with the correlativity of nitrogen balance, show that serum RBP level is sensitivity, the specific index of reflection nutritive disease curative effect.The range of normal value of RBP is male 36-72mg/L, women 22-53mg/L in the blood, requires detection kit that the range of linearity of broad is arranged.
The method that detects RBP at present has unidirectional immunodiffusion RID, enzyme linked immunological absorption EIA, immunoturbidimetry etc.Unidirectional immunodiffusion mainly is manual operations, and test repeatability is relatively poor, and the accuracy of detection is not high, and clinical no longer to have adopted it be quantitative detecting method.The detectability of enzyme linked immunosorbent assay is relatively poor, and time-consuming.At present clinical method of carrying out the normal employing of great amount of samples detection is an immunoturbidimetry.
Can be applied at present and clinically carry out method that a large amount of samples detect immunoturbidimetry is arranged is the turbid and immune scattering turbidimetry of immune transmittance.Immunoturbidimetry is that antigen-antibody combines the dynamic measurement method.Its ultimate principle is: when antigen and antibody reacts in special dilution system and during ratio suitable (general provision antibody excess); The soluble immune complex that forms short in dilution system gathers under the effect of agent (polyglycol etc.); Separate out from liquid phase; Form particulate, make reactant liquor turbidity occur.When AC fixedly the time, the amount of the immune complex of formation increases along with the increase of antigen amount in the sample, and the turbidity of reactant liquor also increases thereupon.Through the turbidity and the contrast of series of standards article of assaying reaction liquid, can calculate the content of antigen in the sample.But when the antigen amount was very low, the antigen-antibody bond was less, was difficult for forming turbidity, so sensitivity is lower.So on the basis of immunoturbidimetry, developed latex enhancing immune than turbid; Its ultimate principle is: antibody is adsorbed on the latex particle of size to fit, uniformity; After antibody and antigen combine, be easy to take place aggegation, because making through light, reduces crosslinked latex particle; Latex particle plays the effect of amplifying signal, can increase sensitivity thus.
Though the kit of panimmunity transmittance purifying method is arranged on the market, because sensitivity is not enough, the shortcoming that all exists is the same reagent RBP in serum sample and the urine specimen simultaneously, and nearly all kit all is the detections that are used for serum sample RBP.Therefore a kind of kit that can detect the RBP in urine specimen and the serum sample of the development of needs is arranged.
Summary of the invention
Therefore, technical purpose of the present invention provides a kind of RBP kit that can detect in urine specimen and the serum sample.
Therefore, first aspect of the present invention relates to a kind of two anti-turbid kit of latex enhancing immune transmittance that encapsulates that detects RBP ELISA, and it comprises reagent 1 and reagent 2 and calibration object; Wherein reagent 1 is to contain the damping fluid with the polymer that promotes cohesion; Reagent 2 is the damping fluids that contain latex-antibody linked thing, and calibration object is the calibration solution of at least 5 variable concentrations of serum matrix or urine matrix, it is characterized in that the antibody in the said reagent 2 is the monoclonal antibody A and the B of pairing; Monoclonal antibody A plays the effect of catching; Monoclonal antibody B plays measuring ability, and preferably, monoclonal antibody A, B are selected from mouse-anti people, the anti-people of chicken, goat anti people's antibody; Preferably, be selected from mouse-anti people antibody.
Preferably, the polymer in the reagent 1 is selected from PEG6000, PEG8000, PEG20000, TWEEN20 and TWEEN80, and concentration range is 0.5-10%.
Preferably, the damping fluid in reagent 1, the reagent 2 is selected from phosphate buffer, Tris, MOPS, and the pH scope of reagent 1 is 6.0-9.0; The pH scope of reagent 2 is 5.5-8.5.
Preferably, monoclonal antibody A consumption is 0.1-0.5mg/ml in the reagent 2, and monoclonal antibody B consumption is 0.1-0.5mg/ml; Preferably; Two monoclonal antibody A and B encapsulate with latex particle respectively, mix then, and monoclonal antibody A and B are anti-human monoclonal antibodies of rabbit or goat-anti human monoclonal antibodies; Preferably, monoclonal antibody A and B are the mouse-anti human monoclonal antibodies.
Preferably, the cross-linking method of preparation reagent 2 is chemical crosslinking or physisorption, and preferably, latex is the granules of polystyrene of carboxyl modified in the chemical crosslinking; Latex is the ordinary polystyrene particle in the physisorption.
Preferably, the particle size range of latex particle is 50-200nm, and preferably, the particle diameter of the latex particle that encapsulates respectively with two monoclonal antibodies is identical or different, and preferably, the concentration range of latex particle is 0.1-0.5% when encapsulating.When the latex particle particle diameter that encapsulates respectively not simultaneously, preferably, the latex particle particle size range 50-150nm that monoclonal antibody A encapsulates, the latex particle particle size range 50-200nm that monoclonal antibody B encapsulates.
Preferably, all add antiseptic in reagent 1, the reagent 2, preferably, Sodium azide and PC300, preferably, consumption is at 0.01-0.3%.
Preferably, adding concentration in the reagent 1 is the EDTA of 10-100mmol/L.
Preferably, the calibration object that is used to detect serum is a serum matrix, and concentration is respectively 130mg/L, 100mg/L, 50mg/L, 25mg/L, 12.5mg/L, 0mg/L, or the content of similar ratio; The calibration object that is used to detect urine is a urine matrix, and concentration is respectively 4mg/L, 2mg/L, 1mg/L, 0.5mg/L, 0.25mg/L, 0mg/L, or the content of similar ratio.
Preferably, serum matrix calibration object and urine matrix calibration object all contain the Sodium azide of 0.01-0.1%, the nystatin of 0.01-0.2%.
Most preferably, the two anti-kit that the turbid kit of latex enhancing immune transmittance is selected from following combination that encapsulates of described detection RBP ELISA:
Kit 1:
Damping fluid MOPS 0.15mol/L pH=6.5,
PEG6000?10％，
NaN
3?0.1％，
EDTA?100mmol/L，
Damping fluid MOPS 0.15mol/L pH=8.5,
Monoclonal antibody A 0.45mg/ml, it is a mouse-anti people antibody, the antigen people urinates the extraction RBP ELISA, tires greater than 1: 5000,
Latex particle A, 104nm, 0.35%, be used for Sheet clonal antibody A,
Monoclonal antibody B 0.5mg/ml, it is a mouse-anti people antibody, the antigen people urinates the extraction RBP ELISA, tires greater than 1: 1000,
Latex particle B, 77nm, 0.5%, be used for Sheet clonal antibody B,
Calibration object A: serum calibration object, concentration are respectively 130,100,50,25,12.5,0mg/L,
Calibration object B: urine calibration object, concentration are respectively 4,2,1,0.5,0.25,0mg/L,
Kit 2:
Damping fluid Tris 0.05mol/L pH=9,
PEG6000?0.56％，
NaN
3?0.1％，
EDTA?10mmol/L，
Damping fluid Tris 0.15mol/L pH=8.0,
Monoclonal antibody A 0.15mg/ml, it is a mouse-anti people antibody, the antigen people urinates the extraction RBP ELISA, tires greater than 1: 5000,
Latex particle A, 159nm, 0.15%, be used for Sheet clonal antibody A,
Monoclonal antibody B 0.12mg/ml, it is a mouse-anti people antibody, the antigen people urinates the extraction RBP ELISA, tires greater than 1: 1000,
Latex particle B, 195nm, 0.11%, be used for Sheet clonal antibody B,
Calibration object A: serum calibration object, concentration are respectively 130,100,50,25,12.5,0mg/L,
Calibration object B: urine calibration object, concentration are respectively 4,2,1,0.5,0.25,0mg/L.
In other words, adopt monoclonal antibody A, the B of pairing to encapsulate respectively on the latex particle among the present invention, the antibody that encapsulates on the latex combines with RBP to be measured, and a plurality of bonds gather together, and formation can change by detected turbidity.Through detecting 6 or more a plurality of calibration object, draw calibration curve.When detecting sample, calculate the concentration of RBP in the sample to be tested through the reaction absorbance.
This kit is owing to adopt two monoclonal antibodies and the latex enhancing; The bond that forms with RBP is bigger than the bond volume that adopts monospecific antibody or do not adopt latex to form; So have higher sensitivity; Can detect the RBP content in the urine accurately, the heavy absorption function of sensitive reflection renal tubule.Through adjustment antibody consumption, this kit can have the range of linearity of broad, is used for detecting serum RBP content simultaneously, the reflection nutrition condition.
The present invention adopts the multiple spot calibration, and having overcome reaction absorbance and concentration is not the directly proportional problem, can guarantee the accuracy of testing result.
The thinking that the present invention adopts double antibody to encapsulate latex particle derives from enzyme mark sandwich method ELISA; (meaning of pairing is meant that monoclonal antibody reacts with antigen respectively to adopt two monoclonal antibodies of matching; Confirm the site of combination; Guarantee that two monoclonal antibodies and antigen reactive binding site are different, such two monoclonal antibodies can be used simultaneously; Monoclonal antibody as capture antibody requires compatibility very good; Detect antibody and require to be applied to signal height in the immune response); Monoclonal antibody A is as capture antibody; Monoclonal antibody B is as detecting antibody, when form two monoclonal antibodies, antigen with and latex three parts that encapsulate when forming bond sensitivity have bigger improvement, do not influence linearity simultaneously.
The present invention adopts immune turbidimetry; The kit that panimmunity transmittance purifying method is arranged on the market; But because sensitivity is not enough, the shortcoming that all exists is same reagent serum sample and a urine specimen simultaneously, and nearly all kit all is the detections that are used for serum sample; The present invention is because two monoclonal antibodies that encapsulate on the latex particle of employing have improved the sensitivity that detects, so overcome this shortcoming.Though in the prior art (" the Retinal-binding protein detection kit that a kind of double antibody is compound ", application number: 20100159837.2, publication number: 101833009A.) also have and relate to the kit that detects RBP ELISA with two kinds of antibody, but pertinent literature does not show the effect that has " pairing " between two kinds of antibody, the method that the method for the document neither adopt latex to strengthen.Simultaneously, the double antibody in the document is a monoclonal antibody and anti-more than, adopts resist to cause cross reaction easily more, so adopt two monoclonal antibodies to help avoid cross reaction among the present invention, improves sensitivity.Though the two reaction principle all is immune responses, the present invention adopts latex to strengthen, and it is high that sensitivity is wanted.Simultaneously, directly with use in the document two anti-when being applied to the immune turbidimetry that latex strengthens effect also be far from the effective of kit of the present invention.Simultaneously, the document is not illustrated the type of sample, but infers it is serum from the value of correlativity sample, because the RBP normal contents in the urine sample is less than 0.7mg/L, so it is wide to detect range of the sample the present invention; Documents is not carried out detailed proof to sensitivity, and that outstanding advantage of the present invention is sensitivity is good.
Description of drawings:
Fig. 1: the linear regression curve of RBP ELISA kit of the present invention.
Fig. 2: RBP ELISA kit of the present invention and external kit detect urinary assay result's correlogram.
Fig. 3: RBP ELISA kit of the present invention and external kit detect determination of serum result's correlogram.
Embodiment
To further specify the present invention through following non-limiting example below, as well known to those skilled in the art, under the situation that does not deviate from spirit of the present invention, can make many modifications to the present invention, such modification also falls into scope of the present invention.
Following experimental technique is conventional method if no special instructions, and employed experiment material all can easily be obtained from commercial company if no special instructions.
Embodiment
Embodiment 1:RBP kit 1 is formed
Damping fluid MOPS 0.15mol/L pH=6.5,
PEG6000?10％，
NaN
3?0.1％，
EDTA?100mmol/L，
Damping fluid MOPS 0.15mol/L pH=8.5,
Monoclonal antibody A 0.45mg/ml (source: mouse-anti people, antigen: the people urinated the extraction RBP ELISA, tires: greater than 1: 5000),
Latex particle A (monoclonal antibody A encapsulates use) 104nm 0.35% (producer: JSR carboxyl amount: 0.238meq/g, concentration: 5%),
Monoclonal antibody B 0.5mg/ml (source: mouse-anti people, antigen: the people urinated the extraction RBP ELISA, tires: greater than 1: 1000),
Latex particle B (monoclonal antibody B encapsulates use) 77nm 0.5% (producer: JSR, carboxyl amount: 0.274meq/g, concentration: 5%),
Calibration object: serum calibration object, concentration are respectively 130,100,50,25,12.5,0mg/L,
Calibration object: urine calibration object, concentration are respectively 4,2,1,0.5,0.25,0mg/L.
Embodiment 2: the sensitivity behaviour evaluation of the application's kit 1
Instrument: Olympus AU400
Table 1: parameter:
Contrast agents:
External certain famous brand name A:
Its main constituent:
Reagent R1:GOOD ' S damping fluid,
Reagent R2: the anti-people RBP of the rabbit that latex encapsulates IgG antibody,
Detect principle:
Latex particle encapsulates how anti-RBP antibody and combines with RBP antigen in the sample, forms immune complex, detects its turbidity at 570nm and changes, and its intensity of variation is directly proportional with RBP content in the sample.
Urine calibration object 5,2.5,1.25,0.625,0.31,0mg/L,
The sensitivity evaluation method:
The sample of the pure article preparation of RBP is diluted, a certain concentration is limited near sensitivity, repeat 10 times and detect water blank and samples of different concentrations, record raw data and reaction absorbance are calculated like following table.
Sensitivity detects data:
(1) embodiment 1 kit 1 sensitivity behaviour evaluation, the result sees table 2-4.
Table 2: kit 1 sensitivity measured value 1
Table 3: the absorbance when kit 1 is measured
Table 4: kit 1 sensitivity statistical appraisal
(2) sensitivity of contrast agents box is estimated, and the result sees table 5-7.
Table 5: contrast agents box sensitivity measured value
Table 6: the absorbance when the contrast agents box is measured
Table 7: contrast agents box sensitivity statistical appraisal
Evaluation criterion: (A-3sd) least concentration greater than 3SDS0 is the sensitivity of this test item.
Conclusion: the sensitivity of the detection kit of detection urine RBP of the present invention is 0.09mg/L, and the sensitivity of contrast agents box is 0.2mg/L, and the sensitivity of kit of the present invention is higher than the sensitivity of contrast agents box far away.
Embodiment 3: the linear properties evaluation of the application's kit 1
Instrument: Olympus 400
Table 8: parameter:
The serum calibration object: concentration is respectively 130,100,50,25,12.5,0mg/L,
The linear evaluation method:
Serum sample is diluted according to 1/10 gradient, be respectively 1/40,1/20,1/10,2/10,3/10,4/10,5/10,6/10,7/10,8/10,9/10,1, each concentration duplicate detection 3 times.With the dilution ratio is the X axle, take the measured value as the mapping of Y axle, and the result sees Fig. 1.
Table 9: linearity test data:
Data are carried out the linear regression mapping, and the result sees accompanying drawing 1 evaluation criterion: detection is compared deviation with theoretical value and should can be accepted ± 10%.
Conclusion: band before concentration of specimens detects after greater than 130mg/L and can occur; The linearity of kit of the present invention is at 7-135mg/L.
The correlativity---urine examination of embodiment 4 the application's kits 1
Reagent, calibration object, parameter all with embodiment 2 in the same; Contrast agents is the above-mentioned external well-known A of producer; Sample is a urine.
Table 10: detect data:
The data that obtained are carried out the correlativity mapping, and the result sees Fig. 2.
Conclusion: negative value appears in contrast agents sometimes when detecting the sample below the 0.15mg/L; The related coefficient of autogamy reagent and import reagent has good correlativity greater than 0.99.
Embodiment 5: the correlativity of the application's kit 1---serum sample
Reagent, calibration object, parameter all with embodiment 4 in the same; Contrast agents is certain external well-known B of producer; Sample is a serum.
External certain famous brand name B:
Main constituent:
Reagent R1: damping fluid,
Reagent R2: the goat anti-human antibody that latex encapsulates,
Calibration object: 150,50,20,10,0mg/L,
Table 11: detect data
The data that obtained are carried out the correlativity mapping, and the result sees Fig. 3.
Conclusion: the related coefficient of autogamy reagent and import reagent has good correlativity greater than 0.99.
Embodiment 6: the component of the application RBP kit 2 and test result thereof
The R1 component:
Damping fluid Tris 0.05mol/L pH=9,
PEG6000?0.56％，
NaN
3?0.1％，
EDTA?10mmol/L，
The R2 component:
Damping fluid Tris 0.15mol/L pH=8.0,
Monoclonal antibody A 0.15mg/ml (source: mouse-anti people, antigen: the people urinated the extraction RBP ELISA, tires: greater than 1: 5000),
Latex particle A (monoclonal antibody A encapsulates use) 159nm 0.15% (producer: JSR carboxyl amount: 0.203meq/g, concentration: 5%),
Monoclonal antibody B 0.12mg/ml (source: mouse-anti people, antigen: the people urinated the extraction RBP ELISA, tires: greater than 1: 1000),
Latex particle B (monoclonal antibody B encapsulates use) 195nm 0.11% (producer: JSR carboxyl amount: 0.128meq/g, concentration: 5%),
Calibration object: serum calibration object, concentration are respectively 130,100,50,25,12.5,0mg/L,
Calibration object: urine calibration object, concentration are respectively 4,2,1,0.5,0.25,0mg/L,
The method of kit 2 according to the foregoing description 2-5 detected, and the result shows that it is all similar at sensitivity, linearity, correlativity aspect of performance and kit 1.
Claims (11)
1. two resisting of detecting RBP ELISA encapsulates the turbid kit of latex enhancing immune transmittance; It comprises reagent 1 and reagent 2 and calibration object, and wherein reagent 1 is to contain the damping fluid with the polymer that promotes cohesion, and reagent 2 is the damping fluids that contain latex-antibody linked thing; Calibration object is the calibration solution of at least 5 variable concentrations of serum matrix or urine matrix; It is characterized in that the antibody in the said reagent 2 is the monoclonal antibody A and the B of pairing, monoclonal antibody A plays the effect of catching, and monoclonal antibody B plays measuring ability; Preferably; Monoclonal antibody A, B are selected from mouse-anti people, the anti-people of chicken, goat anti people's antibody, preferably, are selected from mouse-anti people antibody.
2. the two anti-turbid kit of latex enhancing immune transmittance that encapsulates of detection RBP ELISA according to claim 1; It is characterized in that: the polymer in the reagent 1 is selected from PEG6000, PEG8000, PEG20000, TWEEN20 and TWEEN80, and concentration range is 0.5-10%.
3. the two anti-turbid kit of latex enhancing immune transmittance that encapsulates of detection RBP ELISA according to claim 1 and 2; It is characterized in that: the damping fluid in reagent 1, the reagent 2 is selected from phosphate buffer, Tris, MOPS, and the pH scope of reagent 1 is 6.0-9.0; The pH scope of reagent 2 is 5.5-8.5.
4. two the resisting according to each described detection RBP ELISA of claim 1 to 3 encapsulates the turbid kit of latex enhancing immune transmittance; It is characterized in that: monoclonal antibody A consumption is 0.1-0.5mg/ml in the reagent 2; Monoclonal antibody B consumption is 0.1-0.5mg/ml, and preferably, two monoclonal antibody A and B encapsulate with latex particle respectively; Mix then; Monoclonal antibody A and B are anti-human monoclonal antibodies of rabbit or goat-anti human monoclonal antibodies, and preferably, monoclonal antibody A and B are the mouse-anti human monoclonal antibodies.
5. two the resisting according to each described detection RBP ELISA of claim 1 to 4 encapsulates the turbid kit of latex enhancing immune transmittance; It is characterized in that: the cross-linking method of preparation reagent 2 is chemical crosslinking or physisorption; Preferably, latex is the granules of polystyrene of carboxyl modified in the chemical crosslinking; Latex is the ordinary polystyrene particle in the physisorption.
6. two the resisting according to each described detection RBP ELISA of claim 1 to 4 encapsulates the turbid kit of latex enhancing immune transmittance; It is characterized in that: the particle size range of latex particle is 50-200nm; Preferably; The particle diameter of the latex particle that encapsulates respectively with two monoclonal antibodies is identical or different, and preferably, the concentration range of latex particle is 0.1-0.5% when encapsulating.
7. two the resisting according to each described detection RBP ELISA of claim 1 to 6 encapsulates the turbid kit of latex enhancing immune transmittance; It is characterized in that: all add antiseptic in reagent 1, the reagent 2, preferably, Sodium azide and PC300; Preferably, consumption is at 0.01-0.3%.
8. encapsulate the turbid kit of latex enhancing immune transmittance according to the two anti-of each described detection RBP ELISA of claim 1 to 7, it is characterized in that: adding concentration in the reagent 1 is the EDTA of 10-100mmol/L.
9. two the resisting according to each described detection RBP ELISA of claim 1 to 8 encapsulates the turbid kit of latex enhancing immune transmittance; It is characterized in that: the calibration object that is used to detect serum is a serum matrix, and concentration is respectively 130mg/L, 100mg/L, 50mg/L, 25mg/L, 12.5mg/L, 0mg/L; The calibration object that is used to detect urine is a urine matrix, and concentration is respectively 4mg/L, 2mg/L, 1mg/L, 0.5mg/L, 0.25mg/L, 0mg/L.
10. the two anti-of detection RBP ELISA according to claim 9 encapsulates the turbid kit of latex enhancing immune transmittance, and it is characterized in that: serum matrix calibration object and urine matrix calibration object all contain the Sodium azide of 0.01-0.1%, the nystatin of 0.01-0.2%.
11. encapsulate the turbid kit of latex enhancing immune transmittance according to the two anti-of each described detection RBP ELISA of claim 1 to 9, it is selected from the kit of following combination:
Kit 1:
Reagent 1 component:
Damping fluid MOPS 0.15mol/L pH=6.5,
PEG6000?10％，
NaN
3?0.1％，
EDTA?100mmol/L，
Reagent 2 components:
Damping fluid MOPS 0.15mol/L pH=8.5,
Monoclonal antibody A 0.45mg/ml, it is a mouse-anti people antibody, the antigen people urinates the extraction RBP ELISA, tires greater than 1: 5000,
Latex particle A, 104nm, 0.35%, be used for Sheet clonal antibody A,
Monoclonal antibody B 0.5mg/ml, it is a mouse-anti people antibody, the antigen people urinates the extraction RBP ELISA, tires greater than 1: 1000,
Latex particle B, 77nm, 0.5%, be used for Sheet clonal antibody B,
Calibration object A: serum calibration object, concentration are respectively 130,100,50,25,12.5,0mg/L,
Calibration object B: urine calibration object, concentration are respectively 4,2,1,0.5,0.25,0mg/L,
Kit 2:
Reagent 1 component:
Damping fluid Tris 0.05mol/L pH=9,
PEG6000?0.56％，
NaN
3?0.1％，
EDTA?10mmol/L，
Reagent 2 components:
Damping fluid Tris 0.15mol/L pH=8.0,
Monoclonal antibody A 0.15mg/ml, it is a mouse-anti people antibody, the antigen people urinates the extraction RBP ELISA, tires greater than 1: 5000,
Latex particle A, 159nm, 0.15%, be used for Sheet clonal antibody A,
Monoclonal antibody B 0.12mg/ml, it is a mouse-anti people antibody, the antigen people urinates the extraction RBP ELISA, tires greater than 1: 1000,
Latex particle B, 195nm, 0.11%, be used for Sheet clonal antibody B,
Calibration object A: serum calibration object, concentration are respectively 130,100,50,25,12.5,0mg/L,
Calibration object B: urine calibration object, concentration are respectively 4,2,1,0.5,0.25,0mg/L.
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