CN107607707A - Suppress the bladder chalone C latex enhancing immune of rheumatoid factor interference than turbid kit - Google Patents

Suppress the bladder chalone C latex enhancing immune of rheumatoid factor interference than turbid kit Download PDF

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CN107607707A
CN107607707A CN201710748550.5A CN201710748550A CN107607707A CN 107607707 A CN107607707 A CN 107607707A CN 201710748550 A CN201710748550 A CN 201710748550A CN 107607707 A CN107607707 A CN 107607707A
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reagent
latex
enhancing immune
antibody
kit
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CN107607707B (en
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耿英利
罗湘宇
甘萍萍
黎明
马春霞
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Mike Biological Ltd By Share Ltd
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    • 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
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    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/72—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood pigments, e.g. haemoglobin, bilirubin or other porphyrins; involving occult blood
    • G01N33/721—Haemoglobin
    • 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
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    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
    • 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
    • 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
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    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • 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
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic

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Abstract

The present invention provides a kind of bladder chalone C latex enhancing immune for suppressing rheumatoid factor interference than turbid kit, it includes reagent 1 and reagent 2, the reagent 1 is promotion antigen and the solution of antibody response, the reagent 2 is the latex particle dispersion liquid for being coated with antibody sensitized, and 1 40g/L N HOSu NHSs are included in the reagent 1；0.5 4mmol/L PEG can also be included in the reagent 1.The kit of the present invention can effectively eliminate latex enhancing immune than containing interference of the rheumatoid factor to immune detection result, having high sensitivity, specificity good in turbid sample, quick, measurement result accurately and reliably the characteristics of.

Description

Suppress the bladder chalone C latex enhancing immune of rheumatoid factor interference than turbid kit
Technical field
The present invention relates to external diagnosis reagent field, and in particular to a kind of bladder chalone C glue for suppressing rheumatoid factor interference Breast enhancing Immunoturbidimetric kit.
Background technology
Bladder chalone C (Cystatin, Cys C) is a kind of cystatin, is a kind of small protein, Also referred to as γ-trace of albumin and γ-postglobulin, are widely present in the karyocyte and body fluid of various tissues, are a kind of Low molecule amount, alkaline non-glycated protein, molecular weight 13.3KD, it is made up of, can be had by body is all 122 amino acid residues Nucleus produces, and generation rate is constant.Bladder chalone C in circulation is only eliminated through glomerular filtration, is a kind of reflection glomerulus The endogenous mark of filtration rate change, and in proximal convoluted tubule reabsorption, but blood is not returned to by complete metabolic breakdown after reabsorption Liquid, therefore, its blood level determines by glomerular filtration, and independent of any foeign element, as sex, the age, diet shadow Ring, be a kind of preferable homology mark for reflecting glomerular filtration rate(GFR change.Bladder chalone C in blood circulation can be passed through freely Glomerulus, absorbed and decomposed by epithelial cell in proximal convoluted tubule almost all, only micro discharge in urine.Cys C levels not by appoint What foeign element, such as sex, the influence at age, diet, therefore, Cys C are a kind of reflection glomerular filtration rate(GFR (GFR) changes Preferable endogenous mark.
When impaired renal function, the concentration of Cys C in blood changes and changed with glomerular filtration rate(GFR, during renal failure, kidney Glomerular filtration rate declines, and concentration can increase several times even ten several times to Cys C in blood；If glomerular filtration rate(GFR is normal, and kidney During tubule malfunction, absorptions and catabolism of the Cys C in renal tubule can be hindered, the concentration in urine can increase even to hundred times More than.
Because bladder chalone C concentration is relatively low in serum, therefore its assay method needs higher sensitivity for analysis and specificity.Earliest Bladder chalone C content is measured using simple immunodiffusion method (RID) and enzyme immunoassay (EIA), and both approaches Not only time-consuming, test limit is also poor.Relatively simple and more sensitive radiation, fluorescence and various enzyme immunoassay (EIA)s (RIA, FIA and EIA) Method can improve the reliability of bladder chalone C determining, but minute is still longer, can not routinely apply, more be not used to emergency treatment Measure.Any of the above assay method belongs to heterogeneous assay approach, it is difficult to automate, therefore limits facing extensively for bladder chalone C Bed application.Homogeneous determination method, i.e., the coated antibody on latex particle, with particle during antigen binding are immunized using above-mentioned latex Generation aggegation, the method are easy to automate.Kabanda is equal to the latex particle agglutination of the measure bladder chalone C of report in 1994 Method is a kind of grain count immunoassay.Kyhseandersen reports a kind of immune ratio of particle enhancing transmission equal to the same year Turbid method (particle-enhancedturbidimetric immunoassay, PETIA), makes the time of bladder chalone C determining contract 5min or so is as short as, and can be carried out on automatic biochemistry analyzer, makes it possible that the conventional of bladder chalone C determining is applied.
Latex particle enhancing turbidimetry (PETIA) is that occur in recent years a kind of is relatively stable, accurate body fluid albumen is equal Phase immunoturbidimetry detection method.General principle is the surface-crosslinked antibody in polymer latex microballoon, when being crosslinked with the micro- of antibody Ball with after antigen binding, can rapidly flock together, change the astigmatism performance or light transmission of reaction solution in a short time.And And the change of reaction solution astigmatism performance or light transmission (i.e. absorbance) and the concentration of tested antigen have stronger correlation, The concentration of tested antigen can be reflected in certain limit.PETIA detection methods are that antigen, antibody are carried out in homogeneous reaction system Reaction and the measure of result.After antigen, antibody response, the absorbance of reaction solution is directly determined, ELISA method is eliminated and incubates repeatedly It is time saving and energy saving with regard to that can obtain result with tedious operations step, a few minutes such as board-washing to educate.In addition, nano immune turbidimetry operation step Rapid simplification also accordingly avoid the interference of the extraneous factors such as many manual operation factors and reagent, environment, stability and again Renaturation is all preferable, can more truly reflect the content of measured matter.Although the sensitivity of immunoturbidimetry poorer than ELISA method one A bit, but it is enough to detect the lower limit of many marker proteins in human normal plasma, clinical detection requirement can be fully met.
One limitation of latex enhancing immune turbidimetry conventional detection is susceptible to by being likely to be present in test sample Heterophil antibody caused by disturb, disturbed especially caused by rheumatoid factor.Rheumatoid factor (rheumatoid Factor, RF) be to be found in rheumatoid arthritis (RA) patients serum, be it is a kind of using be denatured IgG as target antigen itself Antibody, it is primarily present in the serum and joint fluid of patient with rheumatoid arthritis, it is a kind of antitypy IgG antibody, category IgM types.It can be combined with IgGFc sections.The B cell clone for producing RF is all there are in patient RA and Yue 50% healthy human body, Under denaturation IgG (or IgG with antigen binding) or Epstein-Barr virus directly act on, RF can be largely synthesized.In the immune class detection examination of business Antibody employed in agent is the IgG antibody-likes of animal origin mostly, and the Fc sections of Fc sections and human IgG have higher homology, Can be identified and combined by RF, so in test substance in detection sample negative, that RF is positive, immunologic function test reagent be possible to by RF captures produce signal, so as to cause false positive results.
At present in the development of existing immunologic function test reagent, the method for reducing rheumatoid factor or the interference of different preferendum is usual Have：(1) interfering immunoglobulin is removed from sample or makes its inactivation.For example use IgG solid-phase adsorbents absorption sample In rheumatoid factor, or use the diluted sample containing 2 mercapto ethanol, with IgM type rheumatoid factor of degrading, but This can make detection complex operation, and it is possible to content or the work of test substance are decreased while removing and disturbing albumen Property.(2) it is mostly IgG, in the market has the sealer of many commercializations using the sealer for reducing interference.But due to rheumatoid The variation of the factor or heterophile antibody, and antibody used in various products are different, at present can be with there has been no a kind of product Effectively eliminate all interference, it usually needs be combined, screened using different product, not only increase workload, very high reagent into This, on the other hand due to using a variety of not clear compositions sealer, also uncontrollably introduce new interference detection results because Element.(3) modification determination antibody is so that it is less susceptible to react with rheumatoid factor.A kind of most currently used method is to detect IgG antibody digestion removes Fc fragments, and complete IgG is substituted with F (ab ') 2, had both retained its activity for identifying binding antibody, and had disappeared again Except Fc interference, the nonspecific reaction of reagent is reduced.But it is anti-to lose a part in the digestion, purifying preparation technology in IgG Body and also having on antibody activity influences to a certain degree, improves reagent cost, it is often more important that and reagent preparation process is further complicated, Impassable obstacle is provided with to control reagent difference between batch, so as to limit the popularization of such reagent clinically.
Existing latex enhancing immune turbidimetry kit, generally comprises reagent 1 and reagent 2, reagent 1 for promote antigen and The buffer solution of antibody response, reagent 2 are the latex particle dispersion liquid for being coated with antibody.
The content of the invention
To solve the above problems, the present invention provides a kind of latex enhancing immune for suppressing rheumatoid factor interference than turbid reagent Box, the kit forms are simple, have extensive versatility than turbid reagent for latex enhancing immune, can effectively eliminate class The interference of the rheumatism factor, improve the specificity and accuracy of testing result.
The present invention provides a kind of bladder chalone C latex enhancing immune for suppressing rheumatoid factor interference than turbid kit, and it is wrapped Reagent 1 and reagent 2 are included, to promote the solution of antigen and antibody response, the reagent 2 resists the reagent 1 to be coated with bladder chalone C The latex particle dispersion liquid of body sensitization, 1-40g/L n-hydroxysuccinimide (NHS) is included in the reagent 1, it is therefore preferable to 5-30g/L, 10-20g/L, 10-30g/L or 20-40g/L n-hydroxysuccinimide.In some embodiments, reagent 1 The amount of middle n-hydroxysuccinimide can be 1g/L, 2g/L, 3g/L, 4g/L, 5g/L, 6g/L, 7g/L, 8g/L, 9g/L, 10g/ L、11g/L、12g/L、13g/L、14g/L、15g/L、16g/L、17g/L、18g/L、19g/L、20g/L、21g/L、22g/L、 23g/L、24g/L、25g/L、26g/L、27g/L、28g/L、29g/L、30g/L、31g/L、32g/L、33g/L、34g/L、35g/ L, 36g/L, 37g/L, 38g/L, 39g/L or 40g/L.
In one embodiment, 0.5mmol/L-4mmol/L PEG is also included in the reagent 1.Polyethylene glycol (PEG), also referred to as poly- (oxirane) (PEO) or polyoxyethylene (POE), the oligomer or polymer of oxirane are referred to.By In the influence of chain length, the polyethylene glycol of different molecular weight often has different physical property (such as viscosity) and different applications, but Most polyethylene glycol chemistry property is similar.PEG can accelerate big immune multiple in latex enhancing immune is than turbid reagent Compound is formed, and is to promote poly- agent, it is possible to increase sensitivity of the latex enhancing immune than turbid reagent.
In one embodiment, the PEG is PEG2000~PEG20000.Such as PEG-2000, PEG-3000, PEG-4000, PEG-6000, PEG8000 and PEG20000.
In one embodiment, NHS concentration is 1-30g/L in the reagent 1.
In one embodiment, the reagent 1 further comprise buffer solution, biological preservative, surfactant and IgM antibody；Include buffer solution, biological preservative, surfactant, sealer and stabilizer with the reagent 2.
Buffer solution can be conventional use of cushioning liquid in this area.In some embodiments, the cushioning liquid MOPS, good's buffer solution, TRIS buffer, phosphate buffer, glycine can be each independently Buffer solution and its any combination.
In some embodiments, the biological preservative can be dichloro acetamide, imidazolidinyl urea, biological preservative ProClin series, potassium sorbate, sodium benzoate, nipagin esters etc. or its any combination.
In some embodiments, it can be TWEEN, SPAN, TRITON, EMULGEN that the surfactant, which can be, Series of surfactants, such as polysorbas20, polysorbate40, TX100 etc..
The effect of sealer is to be combined with-COOH sites of dissociating in latex particle, avoid free carboxyl site Testing result is had undesirable effect, sealer can be various protein in reagent 2 of the present invention, and preferably BSA albumen is as envelope Close agent.
The main function of stabilizer is to protect the activity of antibody and prevent the aggegation of latex particle, sink, while can be with Increase kit in each component stability, extend product shelf life and corkage after usage time.Conventional stabilizer is sugar Class, alcohols, protide and some amino acid etc..In some embodiments, stabilizer is carbohydrate, such as sucrose or marine alga Sugar.
In one embodiment, the reagent 1 includes 25-200mmol/L phosphate buffers, 0.9-120g/L Sodium chloride, 0.5-200mg/L anti-human IgM antibodies, 0.1-5g/L biological preservatives Proclin300 and 1-20g/L surface-active Agent TX100.
In some embodiments, phosphate buffering liquid concentration can be 25-100mmol/L or 100- in reagent 1 200mmol/L.In some embodiments, in reagent 1 amount of sodium chloride be 0.9-30g/L, 30-120g/L, 30-60g/L or 60-80g/L.In some embodiments, the amount of anti-human IgM antibodies is 0.5-50mg/L, 50-100mg/L, 100- in reagent 1 200mg/L or 50-200mg/L.In some embodiments, in reagent 1 amount of biological preservative be 0.1-1g/L, 1-3g/L, Or 1-5g/L.In some embodiments, the amount of the surfactant agent of reagent 1 be 1-3g/L, 1.5-3g/L, 1.5-20g/L or 3-20g/L。
In one embodiment, the reagent 1 include 100mmol/L phosphate buffers, 30g/L sodium chloride, 50mg/L anti-human IgM antibodies, 1g/L biological preservatives Proclin300 and 3g/L surfactant TX100.
In one embodiment, the reagent 2 includes 20-200mmol/L glycine buffers, 0.1-5g/L life Thing preservative Proclin300,1-20g/L surfactant TX100,1-5g/L bovine serum albumin(BSA), 1-200g/L trehaloses With the latex particle of 2-50g/L antibody sensitized.
In some embodiments, glycine buffer concentration can be 20-50mmol/L, 50-200mmol/ in reagent 2 L or 100-200mmol/L.
In some embodiments, the amount of stabilizer is 1-3g/L, 1-150g/L, 3-150g/L or 150- in reagent 2 200g/L.In some embodiments, the amount of biological preservative is 0.1-1g/L, 1-3g/L or 1-5g/L in reagent 2.One In a little embodiments, the amount of surfactant agent is 1-3g/L, 1.5-3g/L, 1.5-20g/L or 3-20g/L in reagent 2.One In a little embodiments, the amount of sealer is 1-2g/L, 1-3g/L, 2-5g/L or 3-5g/L in reagent 2.In some embodiments In, the concentration of the latex particle of antibody sensitized is 0.2-0.5g/L or 0.5-5g/L in reagent 2.
In one embodiment, the reagent 2 includes 50mmol/L glycine buffers, 1g/L biological preservative The antibody of Proclin300,3g/L surfactant TX100,3g/L bovine serum albumin(BSA), 3g/L trehaloses and 0.5g/L causes Quick latex particle.
In one embodiment, it is that animal is anti-human that the latex enhancing immune is more coated than in turbid kit sensitizing latex IgG is polyclonal or monoclonal antibody.The kind of the animal includes rabbit, goat, sheep, chicken and mouse.
The present invention in reagent 1 using adding NHS, after adding NHS, the RF factors are combined with anti-human igg in sample ability Lower, and the binding ability of labelled antibody and determined antigen is unaffected.After adding PEG in reagent 1, the speed of detection is added Degree and sensitivity, and NHS and PEG in both appropriate concentration ranges both have synergy, substantially reduce RF because The influence of son, has more preferable measuring accuracy and the degree of accuracy.In addition, the anti-human IgM antibodies in reagent 1, can with after deactivation The RF factors combine, so as to thoroughly eliminate RF factor pair measurement results interference.Therefore, kit of the invention can improve inspection Survey the specificity and accuracy of result；This other kit forms are simple, have higher sensitivity, have extensive general Property.The present invention compared with the existing technology, has that high sensitivity, specificity are good, quick, measurement result accurately and reliably the characteristics of.
Embodiment
In order that art technology field personnel more fully understand the technical scheme in the application, below in conjunction with embodiment The invention will be further described, it is clear that and described embodiment is only some embodiments of the present application, rather than whole Embodiment.Based on the embodiment in the application, those of ordinary skill in the art are obtained under the premise of creative work is not made The all other embodiment obtained, it should all belong to the scope of the application protection.
Embodiment one:Tested in bladder chalone C determining reagent kit containing different NHS amounts
1. the bladder chalone C determining reagent 1 containing different NHS amounts
Bladder chalone C determining reagent 1-A：200mmol/L phosphate buffers, 120g/L sodium chloride, 200mg/L goat-anti people IgM antibody, 0g/LNHS, biological preservative Proclin300 5g/L, surfactant TX100 2g/L；
Each component is identical with reagent 1-A in bladder chalone C determining reagent 1-B, C, D, E, F, G, H and I, except NHS concentration not Together, they are respectively 1g/L NHS, 5g/L NHS, 10g/L NHS, 20g/L NHS, 30g/L NHS, 40g/L NHS, 45g/L NHS and 50g/L NHS.
2. bladder chalone C determining reagent kit determines reagent 2：200mmol/L glycine buffers, the anti-human bladder chalone Cs of 5.0g/L The latex particle of polyclonal antibody sensitization, 5g/L biological preservatives Proclin300,2g/L surfactant TX100,5g/L ox Seralbumin, 200g/L trehaloses.
3. the bladder chalone C sample preparation containing different rheumatoid factor contents：In the bladder chalone C sterling containing 0.92mg/L Sample in be separately added into different RF, be configured to containing 0,100,200,500,1000IU/mL RF solution.
More than prepare nine kinds of bladder chalone C determining reagents (1-A, 1-B, 1-C, 1-D, 1-E, 1-F, 1-G, 1-H, 1-I, respectively The NHS amounts of reagent are different), to containing 0,100,200,500,1000IU/mL RF bladder chalone C sample detects.
Table 1：Result of the test in bladder chalone C determining reagent kit
Data above is understood：When in reagent 1 without NHS, measurement result substantially raises as RF factor contents increase, by most First 0.92mg/L is increased to 26.43mg/L.NHS is added in reagent 1 can significantly reduce the interference of the RF factors, when NHS contents For 20g/L when, RF factor pairs measurement result influence do not have substantially.But when NHS amounts progressively increase to 45g/L, 50g/L, measure As a result it is obvious relatively low, it is shown as subclutter.
Embodiment two:Bladder chalone C determining reagent kit containing various concentrations PEG4000
1. the bladder chalone C determining reagent containing various concentrations PEG4000
Bladder chalone C determining reagent 1-1：200mmol/L phosphate buffers, 120g/L sodium chloride, 200mg/L goat-anti people IgM antibody, biological preservative Proclin300 5g/L, surfactant TX100 2g/L, 0mmol/LPEG4000
Each component is identical with reagent 1-1 in bladder chalone C determining reagent 1-2,3,4,5,6,7 and 8, except PEG4000 concentration Difference, they be respectively 0.5mmol/LPEG4000,1mmol/LPEG4000,2mmol/LPEG4000,3mmol/LPEG4000, 4mmol/LPEG4000,5mmol/LPEG4000 and 6mmol/LPEG4000.
2. bladder chalone C determining reagent kit determines reagent 2：200mmol/L glycine buffers, the anti-human bladder chalone Cs of 5.0g/L The latex particle of polyclonal antibody sensitization, 5g/L biological preservatives Proclin300,2g/L surfactant TX100,5g/L ox Seralbumin, 200g/L trehaloses.
3. the bladder chalone C sample preparation containing different rheumatoid factor contents：In the bladder chalone C sterling containing 0.92mg/L Sample in be separately added into different RF, be configured to containing 0,100,200,500,1000IU/mL RF solution.
Eight kinds of bladder chalone C determining reagents (1-1,1-2,1-3,1-4,1-5,1-6,1-7,1-8, each reagent prepared above PEG4000 amounts it is different), to containing 0,100,200,500,1000IU/mL RF bladder chalone C sample detects.
Table 2：Result of the test in bladder chalone C determining reagent kit
Data above is understood：When in reagent 1 without PEG4000, measurement result substantially raises as RF factor contents increase, 26.43mg/L is increased to by initial 0.92mg/L.When 0.5-6mmol/LPEG4000 is added in reagent 1, measurement result and nothing When PEG4000 is compared, basic indifference.
Embodiment three:Bladder chalone C determining reagent kit containing various concentrations NHS and PEG4000
1. the bladder chalone C determining reagent containing various concentrations NHS and PEG4000
Bladder chalone C determining reagent 1-A1：200mmol/L phosphate buffers, 120g/L sodium chloride, 200mg/L goat-antis Human IgM antibody, 0g/LNHS, biological preservative Proclin300 5g/L, surfactant TX100 2g/L, 0mmol/L PEG4000
Each component is identical with reagent 1-A1 in bladder chalone C determining reagent 2-B2, C3, D4, E5, F6, G7 and H8, except NHS Different with PEG4000 concentration, they are respectively (1g/L NHS, 0.5mmol/LPEG4000), (5g/L NHS, 1mmol/ LPEG4000), (10g/L NHS, 2mmol/LPEG4000), (20g/L NHS, 3mmol/LPEG4000), (30g/L NHS, 4mmol/LPEG4000), (40g/L NHS, 5mmol/LPEG4000) and (45g/L NHS, 6mmol/LPEG4000).
2. bladder chalone C determining reagent kit determines reagent 2：200mmol/L glycine buffers, the anti-human bladder chalone Cs of 5.0g/L The latex particle of polyclonal antibody sensitization, 5g/L biological preservatives Proclin300,2g/L surfactant TX100,5g/L ox Seralbumin, 200g/L trehaloses.
3. the bladder chalone C sample preparation containing different rheumatoid factor contents：In the bladder chalone C sterling containing 0.92mg/L Sample in be separately added into different RF, be configured to containing 0,100,200,500,1000IU/mL RF solution.
Eight kinds of bladder chalone C determining reagents (1-A1,1-B2,1-C3,1-D4,1-E5,1-F6,1-G7 and the 1- prepared above H8, the NHS amounts of each reagent are different), to containing 0,100,200,500,1000IU/mL RF bladder chalone C sample detects.
Table 3：Result of the test in bladder chalone C determining reagent kit
Data above is understood：When in reagent 1 without NHS and PEG4000, measurement result substantially with RF factor contents increase and Rise, but RF factor pairs measurement result influences not having substantially in 1-B2,1-C3,1-D4,1-E5 and 1-F6 experiment, i.e. NHS It is 0.5mmol/L-4mmol/L for 1-30g/L and PEG4000；In this concentration range, with reagent 1 only have NHS compared with when, When having NHS and PEG4000 simultaneously in reagent 1, measurement result shows, the effect of elimination RF Effects of Factors is more preferable, measuring speed Also faster.
Example IV:Bladder chalone C determining reagent kit containing different molecular weight PEG
1. the bladder chalone C determining reagent containing different molecular weight PEG
Bladder chalone C determining reagent 3-1：200mmol/L phosphate buffers, 120g/L sodium chloride, 200mg/L goat-anti people IgM antibody, 10g/LNHS, biological preservative Proclin300 5g/L, surfactant TX1002g/L,
Each component is identical with reagent 3-1 in bladder chalone C determining reagent 3-2,3,4 and 5, and except PEG species is different, they divide Wei not 1mmol/LPEG2000,1mmol/L PEG4000,1mmol/L PEG6000 and 1mmol/L PEG8000.
2. bladder chalone C determining reagent kit determines reagent 2：200mmol/L glycine buffers, the anti-human bladder chalone Cs of 5.0g/L The latex particle of polyclonal antibody sensitization, 5g/L biological preservatives Proclin300,2g/L surfactant TX100,5g/L ox Seralbumin, 200g/L trehaloses.
3. the bladder chalone C sample preparation containing different rheumatoid factor contents：In the bladder chalone C sterling containing 0.92mg/L Sample in be separately added into different RF, be configured to containing 0,100,200,500,1000IU/mL RF solution.
Five kinds of bladder chalone C determining reagents (3-1,3-2,3-3,3-4 and 3-5, the PEG molecular weight of each reagent prepared above It is different), to containing 0,100,200,500,1000IU/mL RF bladder chalone C sample detects.
Table 4：Result of the test in bladder chalone C determining reagent kit
Data above is understood：When NHS is 10g/L, addition same concentrations 1mmol/L different molecular weight PEG2000, When PEG4000, PEG6000, in the concentration, when compared with there was only NHS in reagent 1, there is 1mmol/L concentration simultaneously in reagent 1 PEG2000, PEG4000, PEG6000 and PEG8000 when, measurement result shows, eliminate RF Effects of Factors effect it is more preferable, survey Measure speed also faster；Basic indifference between different molecular weight PEG2000-8000.
In addition, inventor is also studied for the amount of different material in reagent 2 and reagent 1, find in reagent 2 20-200mmol/L glycine buffers, 0.1-5g/L biological preservative Proclin300,1-20g/L surfactant The latex particle of the antibody sensitized of TX100,1-5g/L bovine serum albumin(BSA), 1-200g/L trehaloses and 0.2-5g/L；And/or 25-200mmol/L phosphate buffers in reagent 1,0.9-120g/L sodium chloride, 0.5-200mg/L goat-antis human IgM antibody, 0.1-5g/L biological preservatives Proclin300 and 1-20g/L surfactant TX100, it is as a result similar with table 1-4.
It should be understood that the present invention disclosed is not limited only to specific method, scheme and the material of description, because these Equal alterable.It will also be understood that purpose of the terminology used here just for the sake of the specific embodiment scheme of description, rather than It is intended to limit the scope of the present invention, the scope of the present invention is limited solely by appended claim.
Those skilled in the art, which will also be appreciated that or be able to confirm that, uses no more than normal experiment, institute herein The many equivalents for the specific embodiment of the invention stated.These equivalents are also contained in appended claim.

Claims (10)

1. suppress the bladder chalone C latex enhancing immune of rheumatoid factor interference than turbid kit, it is characterised in that：Including reagent 1 With reagent 2, for the reagent 1 to promote the solution of antigen and antibody response, the reagent 2 is to be coated with bladder chalone C antibody sensitized Latex particle dispersion liquid, 1-40g/L n-hydroxysuccinimide is included in the reagent 1, it is therefore preferable to 5-30g/L, 10- 20g/L, 10-30g/L or 20-40g/L n-hydroxysuccinimide.
2. the latex enhancing immune according to claim 1 is than turbid kit, it is characterised in that：Also include in the reagent 1 0.5mmol/L-4mmol/L PEG.
3. the latex enhancing immune according to claim 2 is than turbid kit, it is characterised in that：The PEG be PEG2000~ PEG20000。
4. the latex enhancing immune according to claim 3 is than turbid kit, it is characterised in that：NHS concentration in the reagent 1 For 1-30g/L.
5. the latex enhancing immune according to claim 1 is than turbid kit, it is characterised in that：In the reagent 1 further Including buffer solution, biological preservative, surfactant and IgM antibody；With the reagent 2 include buffer solution, biological preservative, Surfactant, sealer and stabilizer.
6. the latex enhancing immune according to claim 5 is than turbid kit, it is characterised in that：The reagent 1 includes 25- 200mmol/L phosphate buffers, 0.9-120g/L sodium chloride, 0.5-200mg/L anti-human IgM antibodies, 0.1-5g/L biologies Preservative Proclin300 and 1-20g/L surfactant TX100.
7. the latex enhancing immune according to claim 6 is than turbid kit, it is characterised in that：The reagent 1 includes 100mmol/L phosphate buffers, 30g/L sodium chloride, 50mg/L anti-human IgM antibodies, 1g/L biological preservatives Proclin300 and 3g/L surfactant TX100.
8. the latex enhancing immune according to claim 5 is than turbid kit, it is characterised in that：The reagent 2 includes 20- 200mmol/L glycine buffers, 0.1-5g/L biological preservative Proclin300,1-20g/L surfactant The latex particle of the antibody sensitized of TX100,1-5g/L bovine serum albumin(BSA), 1-200g/L trehaloses and 0.2-5g/L.
9. the latex enhancing immune according to claim 8 is than turbid kit, it is characterised in that：The reagent 2 includes 50mmol/L glycine buffers, 1g/L biological preservative Proclin300,3g/L surfactant TX100,3g/L ox The latex particle of the antibody sensitized of seralbumin, 3g/L trehaloses and 0.5g/L.
10. the latex enhancing immune according to claim 1 is than turbid kit, it is characterised in that：The latex of the sensitization Coated on grain is that animal anti-human igg is polyclonal or monoclonal antibody.
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