CN101887063A - Human C-reactive protein colloidal gold immunochromatographic assay quantitative test paper - Google Patents
Human C-reactive protein colloidal gold immunochromatographic assay quantitative test paper Download PDFInfo
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Abstract
The invention relates human C-reactive protein (CRP) colloidal gold immunochromatographic assay quantitative test paper, which comprises a sample pad, a bonding pad, a nitrocellulose membrane and absorbent paper mutually lapped on a bottom plate with an adhesive in turn. A colloidal gold-labeled CRP antibody complex is coated on the bonding pad; and a detection line and a quality control line are bonded on the nitrocellulose membrane, and are antibody-enveloped lines. During detection, the samples to be detected contact the lower end of the test paper and are subjected to chromatography along the test paper; if the quality control line and a quantitative line do not develop, the test is ineffective; if only the quality control line develops, the CRP concentration is lower than the lowest detection concentration; and if both the quality control line and the quantitative line become red, the content of the CRP is read through a quantitative detection device. The test paper is characterized in that the quality control line and the quantitative line are respectively arranged from the lower end to the upper end of the test paper along the sample chromatography direction.
Description
Technical field
The invention belongs to the quantitative detection field of human C-reactive albumen, particularly relate to and utilize colloidal gold immunity chromatography to carry out the detection by quantitative of human C-reactive albumen.
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Background technology
C reactive protein (C-reactiveprotein, CRP) be to find in some acute disease patients' serum in nineteen thirty by Tillet and Francis, be ring-type five spheroid albumen, belong to the Oligomeric calbindin, relative molecular mass about 120,000, constitute with non-covalent bond by 5 identical monomers, be that inflammatory lymph factor interleukin-6, il-1, TNF stimulate the liver epithelial cell synthetic.Because the pod membrane C polysaccharide of its energy and streptococcus pneumonia plays precipitation reaction and gains the name.CRP mainly is used as the pointer of bacterial infection and disease activity.When acute inflammation, bacterial infection, large tracts of land disorganization, malignant tumour are arranged in the body, CRP will soon occur and raise, and when curing, be reduced to normal range again soon, this is a kind of acute phase reactive protein (acutephasereactantprotein).
At present, detect CRP clinically and mainly use methods such as immune turbidimetry, simple immunodiffusion method, latex agglutination method.Chinese patent application CN1040438 provides a kind of reagent of fast detecting C-reactive protein, is a kind of latex agglutination method detectable by goat-anti people CRpZgCT, polystyrene latex, glycine buffer, distilled water, Sodium azide preparation.This latex agglutination method is simple fast, but just sxemiquantitative, susceptibility is low, is not suitable for most of quantitative clinical detection that needs.Chinese patent application CN1769894 discloses a kind of kit for determination of high-sensitive C-reactive protein, and comprising reagent R-1 is the styrene latex solution that is combined with goat-anti human C-reactive protein antibodies, and the latex particle diameter is 200-240nm, places suitable damping fluid; Reagent R-2 is the styrene latex solution that is combined with goat-anti human C-reactive protein antibodies, and the latex particle diameter is 30-70nm, places suitable damping fluid.This latex agglutination reaction method has improved sensitivity, can carry out the mensuration of low content level.
Because said method all needs special instrument and equipment, so operating personnel and environment have certain requirement, and also during check fee, loaded down with trivial details, be unfavorable for extensively universal.CRP detection test paper or test card based on immunochromatographic method do not need special instrument and equipment, the transportation that is easy to carry, and easy and simple to handle, chromatography reagent is preserved easily than the detectable of liquid.Wherein, relatively be suitable for clinical fast detecting and method stable be the colloidal gold immunochromatographimethod technology, in clinical medical inspection, occupy one seat with its simple and rapid advantage.China utility model patent CN201096787 relates to a kind of C-reactive protein colored particle diagnose test paper, by utilizing the fast immune chromatographic method, detects the CRP in the clinical samples (whole blood/blood serum) qualitatively.This utility model is made up of base plate, water sucting plate, nitrocellulose filter, colored particle fixed bolster, sample liquid-adsorption layer, MAX line, the base plate middle part is a nitrocellulose filter, a C-reactive protein monoclonal antibody test wire and a sheep anti mouse polyclonal antibody control line are arranged on the nitrocellulose filter, in base plate one end termination is water sucting plate, other end termination is the sample liquid-adsorption layer, the nitrocellulose filter two ends overlap mutually with the colored particle fixed bolster with water sucting plate respectively and are connected, and are pressed with the sample liquid-adsorption layer on the colored particle fixed bolster.But this method can't realize the detection by quantitative of CRP.
Chinese patent application CN101320041 " a kind of colloidal gold method of fast quantitative determination of C-reaction protein and application thereof " provides a kind of colloidal gold immunity percolation method to come the method for detection by quantitative CRP.Reaction principle is the gold-marking immunity spot percolation test of DASP sandwich method, testing sample is evenly mixed in the liquid phase homogeneous medium with immune colloid gold after 40-480 doubly dilutes, change again and be added on the reaction plate, wherein contained, can be the monoclonal antibody of anti-another determinant of determinand fixing on the film or many anti-ly catch specifically with immune colloid gold determinand together, the spot that takes on a red color, CRP concentration is proportional in spot color and luster and the sample.But, because the disclosed detection method of this patent is the colloidal gold immunity percolation method, but not colloidal gold immunity chromatography, so supporting kit need be made up of CRP gold mark liquid dried frozen aquatic products, reaction plate, sample diluting liquid, confining liquid and plastic dropper etc., component is more, causes detecting operation more loaded down with trivial details.Owing to contain gold mark liquid dried frozen aquatic products, this reagent must be before detection equilibrium at room temperature 30 minutes, expended the time of waiting detection, make the promptness of examining report relatively poor.In addition, more crucial is that the sensing range of this kit is relatively limited, occurs the HOOK effect easily in detecting high concentration CRP sample, testing sample must be carried out
SuitableDilution, disclosed as it, dilution 40-480 doubly, but, often when primary detection, can't know what times of this dilution actually, promptly under the situation that the initial concentration of sample can't be known, be easy to generate the not enough situation of enriched sample extension rate, so testing result may be on the low side because of the HOOK effect.Such defective reduces the reliability of this method in detection by quantitative.
Conventional CRP immunochromatography qualitative detection test paper is made up of two lines, is respectively detection line, nature controlling line along the sample trend by lower end to upper end.During mensuration, positive if red decidable all appears in detection line and nature controlling line, if only the nature controlling line appearance is red negative, nature controlling line does not develop the color then for invalid.The detection test paper of Bu Zhiing generally is difficult to qualitative detection like this, realize detection by quantitative on similar detection test paper, must could realize detection by quantitative by special design.
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Summary of the invention
Technical matters to be solved
Technical matters to be solved by this invention provides the human C-reactive protein colloidal gold immunochromatographiassay assay quantitative test paper, to overcome existing human C-reactive protein-colloid gold quantitative measurement technology reagent complexity, complex operation, the relatively slow defective of report speed.
Technical scheme
One of technical scheme of the present invention provides a kind of human C-reactive protein colloidal gold immunochromatographiassay assay quantitative test paper, this detection test paper comprises successively at the sample pad, pad, nitrocellulose filter and the thieving paper that are having mutual overlap joint on the base plate of bonding agent, wherein, scribble the CRP antibody complex of colloid gold label on the described pad, be combined with detection line and nature controlling line on the described nitrocellulose filter, described detection line and nature controlling line are immune antiboidy; During detection, with the following end in contact of testing sample and described detection test paper and along the test paper chromatography, if nature controlling line does not develop the color for invalid, represent that then CRP concentration is less than minimal detectable concentration if having only the nature controlling line colour developing, if nature controlling line and quantitatively line redness all appears, the content by a kind of detection by quantitative device interpretation CRP then; Wherein, be respectively nature controlling line and quantitative line along sample chromatography direction by lower end to the upper end of this detection test paper.
One of optimal way of above-mentioned detection by quantitative test paper is that the CRP antibody complex of described colloid gold label is the mouse-anti CRP monoclonal antibody of mark colloid gold particle.
Two of the optimal way of above-mentioned detection by quantitative test paper is that described nature controlling line and quantitative line coated antibody are CRP antibody.
Two of technical scheme of the present invention provides a kind of human C-reactive protein-colloid gold immunochromatographic method detection by quantitative kit, is made up of the test card, sample diluting liquid and the plastic dropper that comprise above-mentioned detection by quantitative test paper.
Above-mentioned detection by quantitative test paper can become the part of CRP test card, and described test card is made up of above-mentioned detection by quantitative test paper and rigid plastic card.
Three of technical scheme of the present invention provides a kind of preparation method of human C-reactive protein-colloid gold immunochromatographic method quantitative test card, may further comprise the steps successively:
(1) with mouse-anti CRP labeling of monoclonal antibody in the colloid gold particle surface, low-temperature centrifugation, resuspended with borate buffer, centrifugal concentrate obtains collaurum-CRP antibody complex, stored refrigerated;
(2) select for use the plain film of glass fibre as the sample pad material, put it in the PBS sample pad treating fluid that contains BSA, sucrose, PVP and tween and soak dry for standby;
(3) select for use the plain film of glass fibre as the pad material, above-mentioned collaurum-CRP antibody complex is scattered in the Tris buffer diluent that contains sucrose, BSA, PEG2000 and tween again, and be sprayed on the pad and dry;
(4) point sample of nitrocellulose filter: with the diverse location specking antibody of some film instrument, be followed successively by nature controlling line, quantitative line to the upper end, dry from the lower end at nitrocellulose filter;
(5) assembling: thieving paper, nitrocellulose filter, pad, sample pad are attached on the base plate that has bonding agent successively, are cut into test strips, be assembled into test card with the rigid plastic card.
Four of technical scheme of the present invention provides a kind of human C-reactive protein-colloid gold immunochromatography quantitative detecting method, in turn includes the following steps:
(1), utilize the detection by quantitative device to read ratio between detected value and the Quality Control value with the standard antigen of above-mentioned detection detection paper series concentration;
(2) with described ratio to described standard antigen concentration drawing standard curve, typical curve is input in the detection by quantitative device;
(3) contain the testing sample of CRP with above-mentioned detection detection paper, and utilize typical curve in the detection by quantitative device to obtain the CRP concentration value of described testing sample.
Five of technical scheme of the present invention provides a kind of isolated plant of human C-reactive protein-colloid gold immunochromatography detection by quantitative, comprises printer, display screen, keyboard, is fixed in circuit board and optics on the instrument base plate.
Six of technical scheme of the present invention provides the application in the C-reactive protein of a kind of above-mentioned human C-reactive protein colloidal gold immunochromatographiassay assay quantitative test paper in detecting human serum.
Particularly, CRP detection by quantitative test paper of the present invention can be applied to the CRP concentration determination in following several respects:
1. diagnosis of infection and discriminating: because CRP 6～8h after infecting generation promptly begins to raise, 24～48h peaks, and peak value can reach normal hundreds of times, and the hurried decline of its content after infecting elimination can recover normal in one week.And CRP does not have remarkable rising when virus infections, and this discriminating for disease early infection type provides extremely important foundation.
2. determine that antibiotic curative effect: CRP is determining also to have brought into play effect aspect the antibiotic curative effect.A research group of Norway has carried out antibiotic therapy to the 11 routine pharyngitis confirmed cases that infect A type hemolytic streptococcus, and before writing down its treatment simultaneously, CRP level, body temperature and the pharyngalgia of treatment back 1,2,3d alleviate indexs such as degree.The result shows, and compares before the treatment, and 1,2,3 day CRP after the treatment descends 34.1%, 60.1% and 75.3% respectively, and, the degree that CRP reduces and symptom alleviate and treat after fate obviously relevant (P＜0.01).
3. with the relation of angiocardiopathy: the confidential relation that has of low-level CRP(0.1～10mg/L) and angiocardiopathy is the biomarker of cardiovascular inflammation pathology.The super sensitive C-reactive protein (hs-CRP) that the c reactive protein of this level is otherwise known as higher.The individual CRP foundation level and the onset relation of following angiocardiopathy are close.CRP may be than the more effective independently angiocardiopathy of LDL-C prediction index, can increase the prognostic value of the dangerous scoring of lipid examination, metabolic syndrome and Framingham.
4. the monitoring of the state of an illness: CRP inflammatory disease outbreak 6h content promptly raises rapidly, and the duration and the course of disease are suitable, in case disease recovery, CRP content descends rapidly, to the clinical prediction effect that a pioneer is arranged.If CRP continues to raise or go up once again, prompting must be paid attention to.
5. the early diagnosis of newborn child's illness and monitoring: studies show that hs-CRP neonate in early days, especially have abnormal labour history person to carry out conventional sense, and observe dynamic change, significant to early diagnosis, the judgement curative effect of newborn child's illness.The hsCRP average of premature 24h, 72h has significant difference, and the term infant also has similar variation, may point out the immunologic function of neonate 72h and 24h relatively to have marked change, especially aspect the factors such as generation IL-6.The premature raises obviously relevant with infection with term infant's CRP.So think that the neonate is born and survey CRP in the 3d and still have very important meaning.
6. other diseases: hematological system: malignant lymphoma (Hodgkin's disease many), leukaemia etc.; The active stage of collagen disease, anaphylaxis and autoimmune disease (rheumatic fever, SLE, rheumatoid disease etc.), infectious disease, septicaemia and other malignant tumours.
Beneficial effect
The invention provides the quantitative detection test paper of a kind of CRP, the feature of this detection test paper is to be respectively nature controlling line and quantitative line along sample chromatography direction by lower end to upper end.During detection, testing sample is added drop-wise in the well of test card, if nature controlling line does not develop the color for invalid, if redness all appears in nature controlling line and quantitative line, by the detection by quantitative device, utilize based on the optical biosensor of reflectance photometer principle and the content of the direct interpretation CRP of typical curve that presets.CRP detection by quantitative test paper of the prior art realizes that by the colloid gold immune spotting method this method operation steps is loaded down with trivial details relatively, and kit also needs supporting plurality of reagents, and detection efficiency is not high.The detection method of detection test paper of the present invention belongs to the immunochromatography dry method, need not other reagent during detection, get 0.05mL after only needing directly testing sample to be diluted by fixing multiple and splash into well, can go out the result after ten minutes, the operating process simple and fast is compared its detection efficiency of spotting method detection test paper and is obviously improved.This detection test paper can carry out qualitative, quantitative measurement for hospital and individual, and its sensing range is 5-200mg/mL, and when actual sample detected, the curve linear of making was good, R
2Value can reach more than 0.99.All in 0.5%, stability and specificity are all good for error between the actual value of detection by quantitative device readout and actual sample, and the term of validity can reach more than 1 year.
What the difference of the present invention and prior art document CN101320041 was to use in the document is the colloidal gold immunity percolation method, and what the present invention used is colloidal gold immunity chromatography.The method that colloidal gold immunity chromatography detection by quantitative CRP is not arranged at present as yet.Particularly, the matched reagent box of document detection method is made up of CRP gold mark liquid dried frozen aquatic products, reaction plate, sample diluting liquid, confining liquid and plastic dropper etc., and component is more, causes detecting operation loaded down with trivial details.And the supporting kit of the present invention only needs CRP test card, supporting sample diluting liquid and plastic dropper, does not have other complex components, like this, will save several steps during detection, so simple and fast more.Owing to contain gold mark liquid dried frozen aquatic products in the method for the document, must be before detection equilibrium at room temperature 30 minutes, expended the time of waiting detection, make the promptness of examining report relatively poor relatively, detection test paper of the present invention is this defective not.In addition, more crucial is that the sensing range of this kit is limited, occurs the HOOK effect easily in detecting high concentration CRP sample, testing sample must be carried out
SuitableDilution, disclosed as it, dilution 40-480 doubly, but, often when primary detection, can't know what times of this dilution actually, promptly under the situation that the initial concentration of sample can't be known, be easy to generate the not enough situation of enriched sample extension rate, so testing result may be on the low side because of the HOOK effect.Such defective reduces the reliability of this method in detection by quantitative, and detection test paper of the present invention need not to pre-determine extension rate applicable to the sample of high concentration, and test sample is directly by after the fixed proportion dilution, and sensing range is wideer.This detection test paper is applicable to supporting detection by quantitative device, lightly portable, highly sensitive, good stability and exportable testing result, make this detecting operation simple and fast, result accurately and reliably, be easy to recorded and stored.
In this manual, unless have other the explanation, each optimal technical scheme and more preferably the technical characterictic of technical scheme can be combined to form new technical scheme mutually.For concise and to the point purpose, the applicant has omitted the specific descriptions of these combinations in instructions, yet the technical scheme after all these technical characterictic combinations all should be considered to be recorded in this instructions so that clear and definite mode is written.
Embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, as the gold-immunochromatographyreagent reagent for assay operation manual, or the condition of advising according to manufacturer." detection by quantitative device " of the present invention device for utilizing the reflective spectrophotometer principle on the corresponding spectrum of collaurum colour developing, light intensity to be write down, preferably, this device can also be equipped with calculation procedure the ratio of detected value (T) and control value (C) is calculated and record automatically, and the typical curve according to input or setting carries out direct interpretation to the pairing CRP concentration of detected value.
Embodiment 1
The preparation of collaurum-CRP antibody complex
In the 20nm colloid gold particle surface for preparing, low-temperature centrifugation uses borate buffer resuspended to original volume, centrifugal once more 1/10, the 4 ℃ of preservation that is concentrated into original volume with mouse-anti CRP labeling of monoclonal antibody.
Embodiment 2
The preparation of human C-reactive albumen (CRP) immunochromatographydetection detection card
(1) preparation of sample pad: select for use the plain film of glass fibre as the sample pad material, put it in the sample pad treating fluid (PBS contains BSA, sucrose, PVP, tween) and soak 37 ℃ of dry for standby.
(2) preparation of pad: select for use the plain film of glass fibre as the pad material, the collaurum-CRP antibody complex of above-mentioned preparation is scattered in the collaurum dilution again, and (Tris buffer solution contains sucrose, BSA, PEG20000, tween) in, and be sprayed on the pad 37 ℃ of oven dry.
(3) point sample of nitrocellulose filter (NC film):, be followed successively by nature controlling line, quantitative line to the upper end from the lower end, 37 ℃ of oven dry with the diverse location specking corresponding antibody of some film instrument at the NC film.
(4) assembling: thieving paper, NC film, pad, sample pad are attached on the base plate that has bonding agent successively, and are cut into the wide test strips of 4cm, assembling finished product detection card.
Embodiment 3
The detection by quantitative of people CRP serum
(1) CRP antigen standard items is disposed the series concentration standard items with normal human serum as dilution: 0mg/mL, 5mg/mL, 10mg/mL, 20mg/mL, 30mg/mL, 50mg/mL, 80mg/mL, 100mg/mL, 120mg/mL, 160mg/mL, 200mg/mL.
(2) as above the standard items of each concentration dilute by fixed proportion, get in the well that 0.05mL is added drop-wise to test card, read the ratio of quantitative line and nature controlling line signal intensity behind the 10min by the detection by quantitative device, be that the T/C value (measure 3 times with 3 test card respectively by each sample, average), draw corresponding standard curve.
(3) utilize above-mentioned typical curve to detect clinical sample, with a among the typical curve y=ax+b that draws, b value input detection by quantitative device can directly draw the content of CRP in the actual sample when measuring actual sample.
Embodiment 4
The detection by quantitative of clinical CRP sample
(1) preparation of quantitative criterion curve: that gets several concentration contains the CRP human serum as standard antigen, and guarantees that each concentration has the even distribution of gradient in monitoring range.Detect the serum antigen of each concentration respectively with above-mentioned CRP gold mark test card, detect the T/C value of the test card of each concentration respectively with supporting detection by quantitative device, each detects three times.Three groups of T/C values that detect are averaged respectively, T/C mean value is figure, obtain a curve with the concentration of described blood serum sample.The result shows, utilizes the prepared typical curve of test card of the present invention linear good, does not occur the HOOK effect in monitoring range.R
2Reach more than 0.99.
(2) utilize typical curve that clinical CRP blood serum sample is carried out detection by quantitative:, to read the T/C value with the detection by quantitative device with above-mentioned test card examination criteria antigen.The result draws out typical curve with gained, obtains corresponding a, the b value.With a, the b value is input in the detection by quantitative device.Getting 150 parts contains the clinical antigen samples of CRP human serum and makes its concentration within the 5-200mg/mL scope.Detect the sample of each concentration respectively with the detection by quantitative device, the CRP concentration numerical value that record detection by quantitative device provides.The CRP concentration value that the detection by quantitative device obtains is compared with the concentration value of typical curve, and the result shows that two groups of numerical value are consistent with each other.With SPSS13.0 two groups of numerical value are carried out correlation analysis, the result is two groups of numerical value no significant differences.Two groups of concentration numerical value are carried out error analysis, the error that has obtained measured concentration and actual concentrations all 0.5% with interior result.
(3) test card stability test: the test card for preparing is placed in 37 ℃ of environment carries out accelerated test.Take out test card after 7 days, and detect the concentration of CRP in the actual sample with it.Measured concentration and actual sample concentration numerical value are carried out error analysis, obtain the CV value 5% with interior result.
Claims (9)
1. human C-reactive protein colloidal gold immunochromatographiassay assay quantitative test paper, this detection test paper comprises successively at the sample pad, pad, nitrocellulose filter and the thieving paper that are having mutual overlap joint on the base plate of bonding agent, wherein, scribble the CRP antibody complex of colloid gold label on the described pad, be combined with quantitative line and nature controlling line on the described nitrocellulose filter, described quantitative line and nature controlling line CRP antibody sandwich; During detection, with the following end in contact of testing sample and described detection test paper and along the test paper chromatography, if nature controlling line does not develop the color for invalid, represent that then CRP concentration is less than minimal detectable concentration if having only the nature controlling line colour developing, if nature controlling line and quantitatively line redness all appears, the content by a kind of detection by quantitative device interpretation CRP then; It is characterized in that, be respectively nature controlling line and quantitative line by lower end to the upper end of this detection test paper along sample chromatography direction.
2. detection by quantitative test paper according to claim 1 is characterized in that, the CRP antibody complex of described colloid gold label is the mouse-anti people CRP monoclonal antibody of mark colloid gold particle.
3. detection by quantitative test paper according to claim 1 is characterized in that, described coated antibody is a CRP antibody.
4. a human C-reactive protein-colloid gold immunochromatographic method detection by quantitative kit is made up of the test card that comprises the described detection by quantitative test paper of claim 1, sample diluting liquid and plastic dropper.
5. kit according to claim 4 is characterized in that, described test card is made up of described detection by quantitative test paper of claim 1 and rigid plastic card.
6. the preparation method of human C-reactive protein-colloid gold immunochromatographic method quantitative test card may further comprise the steps successively:
(1) with mouse-anti people CRP labeling of monoclonal antibody in the colloid gold particle surface, low-temperature centrifugation, resuspended with borate buffer, centrifugal concentrate obtains collaurum-CRP antibody complex, stored refrigerated;
(2) select for use the plain film of glass fibre as the sample pad material, put it in the PBS sample pad treating fluid that contains BSA, sucrose, PVP and tween and soak dry for standby;
(3) select for use the plain film of glass fibre as the pad material, above-mentioned collaurum-CRP antibody complex is scattered in the Tris buffer diluent that contains sucrose, BSA, PEG2000 and tween again, and be sprayed on the pad and dry;
(4) point sample of nitrocellulose filter: with the diverse location specking antibody of some film instrument, be followed successively by nature controlling line, quantitative line to the upper end, dry from the lower end at nitrocellulose filter;
(5) assembling: thieving paper, nitrocellulose filter, pad, sample pad are attached on the base plate that has bonding agent successively, are cut into test strips, be assembled into test card with the rigid plastic card.
7. human C-reactive protein-colloid gold immunochromatography quantitative detecting method in turn includes the following steps:
(1), utilize the detection by quantitative device to read ratio between detected value and the Quality Control value with the standard antigen of claim 1 described detection detection paper series concentration;
(2) with described ratio to described standard antigen concentration drawing standard curve, typical curve is input in the detection by quantitative device;
(3) contain the testing sample of CRP with the described detection detection paper of claim 1, and utilize typical curve in the detection by quantitative device to obtain the CRP concentration value of described testing sample.
8. the isolated plant of a human C-reactive protein-colloid gold immunochromatography detection by quantitative comprises printer, display screen, keyboard, is fixed in circuit board and optics on the instrument base plate.
9. the described human C-reactive protein colloidal gold immunochromatographiassay assay quantitative test paper of claim 1 application in the C-reactive protein in detecting human serum.
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