CN204287198U - Omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper - Google Patents
Omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper Download PDFInfo
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- CN204287198U CN204287198U CN201420645294.9U CN201420645294U CN204287198U CN 204287198 U CN204287198 U CN 204287198U CN 201420645294 U CN201420645294 U CN 201420645294U CN 204287198 U CN204287198 U CN 204287198U
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Abstract
The utility model discloses a kind of omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper, comprise base plate, sample pad, fluorescent marker pad, cellulose nitrate and adsorptive pads; Described test strips is overlapped successively to be pasted onto on base plate formed by sample pad, fluorescent marker pad, cellulose nitrate, adsorptive pads.Described fluorescent marker pad contains marked by streptavidin fluorescin and biotin labeled omnidistance C reactive protein monoclonal antibody; Described nitrocellulose membrane has detection line and nature controlling line, detection line is by omnidistance C reactive protein monoclonal antibody, and it has different identification epi-positions from above-mentioned biotin labeled omnidistance C reactive protein monoclonal antibody.The utility model, compared with the method for the omnidistance C reactive protein of detection common at present (as: chemoluminescence method/Immune-enhancing effect turbidimetry/colloidal gold method), not only substantially reduces detection time, also improves detection sensitivity simultaneously.
Description
Technical field
The utility model belongs to field of immunodetection, is specifically related to the omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper of a kind of high sensitivity.
Background technology
Mankind's c reactive protein (CRP) is that plasma concentration is quick when infection and tissue damage, the main acute phase protein sharply raised.C reactive protein can activating complement and strengthen cytophagous engulfing and play opsonic action, thus removes the invasion pathogenic microorganism of body and damage, and downright bad, the histocyte of apoptosis, plays important protective effect in the innate immunity process of body.Research about C reactive protein has had the history of more than 70 year, traditional view thinks that C reactive protein is a kind of non-specific marker of inflammation, but the research of nearly ten years discloses CRP and participated in the angiocardiopathy such as inflammation and atherosclerotic directly, and be the most strong indication factor of angiocardiopathy and risk factor.
The detection of C reactive protein was widely used in clinical as the non-specific markers of inflammation and tissue damage before the eighties.But due to the detection method comparatively backwardness of past C reactive protein, false positive and false negative are very high, have impact on its value clinically, and ignored by clinical gradually.In recent years, due to the renewal of detection technique, quick, the easy and reliable method measuring C reactive protein is set up rapidly, and C reactive protein is increased greatly in clinical practice field.
The common method of current detection C reactive protein has:
1 simple immunodiffusion method (be called for short and singly expand method)
Single expansion method is a kind of antigen-antibody precipitation test of classics, its ultimate principle is in agaropectin, be mixed into a certain amount of antibody, make determined antigen solution from local to free diffusing in agar, in precipitation zone, form visible precipitation ring, the size of deposit ring diameter or area is relevant to antigen amount.Single expansion method has specificity high as the method for simple and easy antigen quantify, reproducible, simple to operate, cheap, does not need the advantages such as specific apparatus detection.Thus, apply more at some small-middle hospitals.But the maximum shortcoming of this method is when antigen excess, there is not precipitation in reaction system, during CRP excessive concentration, there will be higher false negative.Therefore, when there is not precipitation ring with simple immunodiffusion method detection CRP, recheck after sample must be diluted, to avoid failing to pinpoint a disease in diagnosis.In addition, because the susceptibility of this method is poor, its widespread use is clinically constrained.
2 Latex Agglutination
Latex Agglutination is clinical comparatively conventional serological method, belongs to indirectly agglutination test.The anti-human CRP antibody sensitized of emulsion reagent purifying, can and patients serum in CRP there is specific reaction, present agglutinating particle clearly in several minutes, occur that aggegation person is for positive, do not occur that aggegation person is for feminine gender.The method is simple to operate, quick, and susceptibility, specificity are higher.But be subject to the interference of the factor such as complement, rheumatoid factor (RF), produce false positive results.Therefore, in order to improve the accuracy of result, tackling sample to be measured during detection and carrying out pre-service, to remove disturbing factor.
3 rate nephelometry
Rate nephelometry is the skeptophylaxis precipitation method that Sternberg creates for 1977, is measure the content that the scattering degree of solution to light carrys out antigen in judgement sample.The light of certain wavelength irradiates along transverse axis, and encountering short grained immune complex can cause light scattering, and scattering strength is directly proportional to the content of antigenantibody complex.
This method is a kind of dynamic method of antigen-antibody binding reaction, can measure the content of antigen in sample quickly and accurately, and can on multiple automatic detecting instrument measurement result.Current application more widely self-reacting device is the immunoanalyzer that Backman-Coulter company produces.Rate nephelometry is clinically as CRP conventional sense means.There is research display: rate nephelometry is all better than single expansion method in detection time, method of operating, result degree of accuracy, repeatability, sensitivity.Sensitivity, accuracy also have significant difference (P < 0.05) compared with Latex Agglutination, apparently higher than Latex Agglutination.
4 Immunity transmission turbidity
Immunity transmission turbidity is that the method detecting CRP is commonly used in laboratory, and current many producers also all use this method, and the CRP of Yingke Xinchuang (Ximen) Sci. & Tech. Co., Ltd. is exactly this method.Immunity transmission turbidity is also a kind of immunoprecipitation assay of trace.Itself and rate nephelometry are unlike the content reflecting determined antigen with the light quantity measured through solution.When light therethrough reaction system, the antigenantibody complex in solution can be absorbed light and be reflected, and transmitted light is reduced.Immune complex is more, and the light of absorption is more, and transmitted light is fewer, and the available absorbance of this change represents.If antibody amount is fixed, survey absorbance and be directly proportional to the amount of immune complex, be also directly proportional to the amount of determined antigen.Compare with the antigen standard of a series of concentration known, namely can measure examined object content.
5 Latex-enhanced immunoturbidimetric assays
In above-mentioned turbidimetry, the susceptibility aspect detecting CRP is still not ideal enough.This just needs to set up detection CRP method more responsive, more accurate, still have high accuracy when Serum CRP concentrations is lower.So people improve detection method and upgrade, found Latex-enhanced immunoturbidimetric assay.
Latex-enhanced immunoturbidimetric assay ultimate principle is first adsorbed on by antibody on a kind of latex particle, and when running into corresponding antigen, antigen-antibody combines and occurs latex agglutination.The size of single latex particle within lambda1-wavelength, light-transmissive.When two or more latex particle agglutination, can hinder light therethrough, transmitted light is reduced, its minimizing degree is directly proportional to the degree of latex agglutination, is also directly proportional to antigen amount.The method measures the quick detection method of High-sensitivity Creactive protein (Hs-CRP) a kind of novel height, has the advantages such as quick, convenient, accurate.
6 immunolabelling techniques
The immune labeled method that immunolabelling technique is used for CRP mensuration has radioimmunology, enzyme immunoassay, gold-marking immunity method, fluorescent mark immunity method etc.Due to radioimmunology, to there is the radioactive isotope half life period short, and the shortcomings such as radioactive contamination is not easily preserved, poor stability, have inconvenience in using, especially the widespread use of enzyme immunoassay, and the method is seldom adopted now clinically.At present, clinical practice comparatively multi-method be based on the enzyme immunolabelling technique of enzyme linked immunosorbent assay (ELISA).The method is combined the effect of substrate efficient catalytic with enzyme the specificity of antigen-antibody reaction, according to color change after zymolyte, carries out quantitative measurement and analysis, and calculate CRP content with standard curve control by microplate reader to detection sample.ELISA method has susceptibility, the specificity of height, and its reagent is more stable, no radioactivity pollute.Especially the application of commercially available reagent box and robotization microplate reader, becomes the detection means being applicable to inspection department at different levels.Meanwhile, be also measure one of conventional method of patients serum Hs-CRP.
Carry out the detection of quantitative CRP by Gold standard and fluorescent mark immunity chromatography, in detection sensitivity and test agility, obtain reasonable unification.Advantage is that detection sample consumption is few, and detect fast, can detect together with routine blood test, specificity is high, in clinical detection, also demonstrate good application prospect.Current homemade goods and Korea S, Canadian imported product have had certain application at home.
Biotin-avidin system (biotinavidin system, BAS) is a kind of new bio reaction amplification system.Along with the appearance of various biotin derivative, BAS is widely used in each field of medical science very soon.This system is applied to SABC, enzyme linked immunological, fluorescence immunoassay, in the detection techniques such as radio-immunity, the susceptibility of above technical method, specificity and stability can be improved significantly, make method easier, contribute to clinical quick diagnosis, and be beneficial to and carry out extensive epidemiology survey, become the immunoreactive powerful of research.Because BAS detection system economy is quick, pollute without radiating matter again, do not need complex instrument, fully show the great potential of this system and the possibility of application.
Biotin-avidin system Cleaning Principle: BAS is on the basis of conventional ELISA principle, in conjunction with the height amplification between biotin and Avidin, and a kind of detection system set up.Avidin is a kind of alkaline glycoprotein extracted in ovalbumin, and molecular weight is 68kDa, is made up of 4 subunits, have very high affinity (binding constant is up to 1015M-1) to biology.Biotin is very easily combined with covalent bond with protein (as antibody etc.).Like this, the Avidin molecule combining enzyme and the biotin molecule being combined with specific antibody produce and react, both served multistage amplification, the colour generation due to the catalytic action of enzyme when running into corresponding substrate, reaches the object detecting unknown antigen (or antibody) molecule again.Streptavidin is and the affine a kind of protein have similar biological properties, in Streptavidin molecule the same as Avidin, every bar peptide chain can in conjunction with a biotin, because nearly all material for marking all can combine with Avidin or streptavidin.Utilize biotin-avidin system to develop Procalcitonin detection examination fast diagnosis reagent and there is no report.
Utility model content
The purpose of this utility model, be to provide a kind of omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper, it is based on double antibody sandwich method, fluorescence immunoassay lateral chromatography and biotin-Streptavidin amplification system, provides high sensitivity omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper.When using the utility model, improve detection sensitivity and detect stability, reducing non-specific binding, be not more than 10 minutes detection time, substantially increase diagnosis efficiency.
The solution that the utility model solves its technical matters is: omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper, it comprises base plate, described base plate is provided with sample pad successively, fluorescent marker pad, nitrocellulose filter and adsorptive pads, described adsorptive pads and fluorescent marker pad are overlapping to be respectively pressed in behind the two ends of nitrocellulose filter in the formation detection zone, surface of nitrocellulose filter, described sample pad is overlapping to be pressed on fluorescent marker pad, described fluorescent marker pad is fixed with the fluorescin of biotin labeled omnidistance C reactive protein monoclonal antibody and marked by streptavidin, nitrocellulose filter in described detection zone is fixed with the nature controlling line of detection line and the sheep anti-mouse igg polyclonal antibody formation identifying that the monoclonal antibody of omnidistance C reactive protein another one epi-position is formed.
As the further improvement of technique scheme, described fluorescent marker pad comprises the first stacked fluorescent marker pad and the second fluorescent marker pad, one end pad of described first fluorescent marker pad in the below of sample pad, described second overlapping one end being pressed in nitrocellulose filter of fluorescent marker pad.
As the further improvement of technique scheme, in described first fluorescent marker pad, the one end of inserting below sample pad is provided with positioning strip, and the lower surface of described sample pad is provided with the locating slot that can hold positioning strip.
As the further improvement of technique scheme, also comprise and getting stuck, in described base plate, sample pad, fluorescent marker pad, nitrocellulose filter and adsorptive pads are all placed in and get stuck, the position described shell face of getting stuck corresponding to nitrocellulose membrane is provided with view window, and position shell face of getting stuck corresponding to sample pad is provided with well.
As the further improvement of technique scheme, described view window is slot.
As the further improvement of technique scheme, described fluorescin can be the one in green fluorescent protein, phycobniliprotein.
As the further improvement of technique scheme, described sample pad is glass fibre component dry after surfactant damping fluid immersion treatment.
As the further improvement of technique scheme, described base plate is polystyrene component or tygon component.
As the further improvement of technique scheme, described in get stuck and comprise plastics upper casing and plastics lower casing, described plastics upper casing fastens to be formed after on plastics lower casing and gets stuck.
Compared with prior art, tool has the following advantages the utility model:
(1) the utility model is using fluorescin as label, and this label has good stability, and is conducive to improving detecting stability.
(2) the utility model is by utilizing " Streptavidin-biotin amplification system ", improves detection sensitivity, reduces non-specific binding, is conducive to improving kit performance.
(3) detection time utilizing the utility model to carry out Procalcitonin is not more than 10 minutes, and the detection range of linearity is 0.10ng/ml ~ 100.00ng/ml, drastically increases detection efficiency.
(4) the utility model carries out interpretation by fluorescence immunity analyzer to result, can realize robotization, reduces the impact of subjective factor, provides convenient, quick, reliable diagnostic result.
(5) novel the getting stuck of this use is provided with the well that sample enters and the view window supplying observed result, according to analytical instrument result of determination, accurately and reliably.
(6) the utility model is easy to make, and volume is little, be easy to carry.
(7) the utility model testing cost is lower.
(8) the utility model can be mass, and is applicable to clinical quick diagnosis and on-the-spot quick diagnosis; Be easy to preserve, be conducive to grass-roots unit and promote.
Accompanying drawing explanation
In order to be illustrated more clearly in the technical scheme in the utility model embodiment, below the accompanying drawing used required in describing embodiment is briefly described.Obviously, described accompanying drawing is a part of embodiment of the present utility model, instead of whole embodiment, and those skilled in the art, under the prerequisite not paying creative work, can also obtain other design proposals and accompanying drawing according to these accompanying drawings.
Fig. 1 is the structural representation of the utility model embodiment one;
Fig. 2 is the structural representation of the utility model embodiment two;
Fig. 3 is the structural representation of the utility model embodiment three;
Fig. 4 is the structural representation of the utility model embodiment four.
Embodiment
Be clearly and completely described below with reference to embodiment and the accompanying drawing technique effect to design of the present utility model, concrete structure and generation, to understand the purpose of this utility model, characteristic sum effect fully.Obviously; described embodiment is a part of embodiment of the present utility model, instead of whole embodiment, based on embodiment of the present utility model; other embodiments that those skilled in the art obtains under the prerequisite not paying creative work, all belong to the scope of the utility model protection.In addition, all connection/annexations mentioned in literary composition, not singly refer to that component directly connects, and refer to and according to concrete performance, can connect auxiliary by adding or reducing, and form more excellent draw bail.
With reference to Fig. 1, omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper, it comprises base plate 5, described base plate 5 is provided with sample pad 1 successively, fluorescent marker pad 2, nitrocellulose filter 3 and adsorptive pads 4, described adsorptive pads 4 and fluorescent marker pad 2 are overlapping to be respectively pressed in behind the two ends of nitrocellulose filter 3 in the formation detection zone, surface of nitrocellulose filter 3, fluorescent marker pad 2 is glass fibre membrane, described sample pad 1 is overlapping to be pressed on fluorescent marker pad 2, the fluorescin described fluorescent marker pad 2 being fixed with biotin labeled omnidistance C reactive protein monoclonal antibody (concentration 0.3 ~ 1.5mg/mL) and marked by streptavidin (or Avidin) (uses excitation wavelength 530nm, wavelength of transmitted light 570nm, concentration 0.1 ~ 1.0mg/mL), nitrocellulose filter 3 in described detection zone is fixed with the nature controlling line C (concentration 0.2 ~ 2.0mg/mL) of detection line T (concentration 0.5 ~ 3mg/mL) and the sheep anti-mouse igg polyclonal antibody formation identifying that the monoclonal antibody of omnidistance C reactive protein another one epi-position is formed, nature controlling line C is used for the validity of test strip.
The fluorescin that biotin labeled C reactive protein monoclonal antibody and Streptavidin (or Avidin) mark is fixed on fluorescent marker pad 2, the monoclonal antibody and sheep anti-mouse igg polyclonal antibody that have identification C reactive protein another one epi-position are fixed on nitrocellulose filter as detection line T and nature controlling line C.When testing sample is added to after in sample pad 1, moved forward by chromatography effect, in sample, on C reactive protein and fluorescent marker pad 2, the C reactive protein antibody (Mab-CRP*Fluoro) of combined with fluorescent label (fluorescin) reacts and forms compound CRP-Mab-CRP*Fluoro, compound is reacted when continuing to be advanced past CRP antibody (detection line) nitrocellulose membrane wrapping quilt under chromatography effect, CRP antibody capture formation compound (Mab-CRP-CRP-Mab-CRP*Fluoro) (detection line) that reaction compound is coated, the reaction signal of detection line is read by fluorescence immunity analyzer, under the effect of excitation source, fluorescent material launches the fluorescence signal of specific wavelength, fluorescence immunity analyzer captures fluorescence signal, the typical curve transformed by signal and set is converted into quantitative value automatically, calculate the concentration of C reactive protein in sample, obtain C reactive protein testing result.
As the further improvement of technique scheme, with reference to Fig. 3, described fluorescent marker pad 2 comprises the first stacked fluorescent marker pad 20 and the second fluorescent marker pad 21, one end pad of described first fluorescent marker pad 20 in the below of sample pad 1, described second overlapping one end being pressed in nitrocellulose filter 3 of fluorescent marker pad 21.The layering of fluorescent marker pad 2 is arranged, and is convenient to fluorescent marker pad 2 to be arranged between sample pad 1 and nitrocellulose filter 3.
As the further improvement of technique scheme, be provided with positioning strip 22 with reference to one end of inserting below sample pad in the first fluorescent marker pad 20 described in Fig. 3, the lower surface of described sample pad 1 is provided with the locating slot that can hold positioning strip 22.The installation be convenient between fluorescent marker pad 2 and sample pad 1 by positioning strip 22 is fixed.
As the further improvement of technique scheme, with reference to Fig. 2 and Fig. 4, also comprise and getting stuck, described base plate 5, sample pad 1, fluorescent marker pad 2, nitrocellulose filter 3 and adsorptive pads 4 are all placed in and get stuck in 6, the position described 6 shell faces of getting stuck corresponding to nitrocellulose membrane 3 is provided with view window 8, and the position 6 shell faces of getting stuck corresponding to sample pad 1 is provided with well 7.
As the further improvement of technique scheme, described view window 8 is slot.
As the further improvement of technique scheme, described fluorescin can be the one in green fluorescent protein, phycobniliprotein.
As the further improvement of technique scheme, described sample pad 1 is glass fibre component dry after surfactant damping fluid immersion treatment.Sample pad is made up of glass fibre, through surfactant damping fluid immersion treatment, uses after dry.The cutting width of described sample pad 1 is 0.3 ~ 0.5cm.
As the further improvement of technique scheme, described base plate 5 is polystyrene component or tygon component.
As the further improvement of technique scheme, described in get stuck and 6 comprise plastics upper casing 60 and plastics lower casing 61, described plastics upper casing 61 fastens to be formed after on plastics lower casing 61 and gets stuck 6.
More than that better embodiment of the present utility model is illustrated, but the invention is not limited to described embodiment, those of ordinary skill in the art also can make all equivalent modifications or replacement under the prerequisite without prejudice to the utility model spirit, and these equivalent modification or replacement are all included in the application's claim limited range.
Claims (9)
1. omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper, it is characterized in that: it comprises base plate, described base plate is provided with sample pad successively, fluorescent marker pad, nitrocellulose filter and adsorptive pads, described adsorptive pads and fluorescent marker pad are overlapping to be respectively pressed in behind the two ends of nitrocellulose filter in the formation detection zone, surface of nitrocellulose filter, described sample pad is overlapping to be pressed on fluorescent marker pad, described fluorescent marker pad is fixed with the fluorescin of biotin labeled omnidistance C reactive protein monoclonal antibody and marked by streptavidin, nitrocellulose filter in described detection zone is fixed with the nature controlling line of detection line and the sheep anti-mouse igg polyclonal antibody formation identifying that the monoclonal antibody of omnidistance C reactive protein another one epi-position is formed.
2. omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper according to claim 1, it is characterized in that: described fluorescent marker pad comprises the first stacked fluorescent marker pad and the second fluorescent marker pad, one end pad of described first fluorescent marker pad in the below of sample pad, described second overlapping one end being pressed in nitrocellulose filter of fluorescent marker pad.
3. omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper according to claim 2, it is characterized in that: in described first fluorescent marker pad, the one end of inserting below sample pad is provided with positioning strip, and the lower surface of described sample pad is provided with the locating slot that can hold positioning strip.
4. the omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper according to any one of claims 1 to 3, it is characterized in that: also comprise and getting stuck, in described base plate, sample pad, fluorescent marker pad, nitrocellulose filter and adsorptive pads are all placed in and get stuck, the position described shell face of getting stuck corresponding to nitrocellulose membrane is provided with view window, and position shell face of getting stuck corresponding to sample pad is provided with well.
5. omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper according to claim 4, is characterized in that: described view window is slot.
6. omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper according to claim 4, is characterized in that: described fluorescin can be the one in green fluorescent protein, phycobniliprotein.
7. omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper according to claim 4, is characterized in that: described sample pad is glass fibre component dry after surfactant damping fluid immersion treatment.
8. omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper according to claim 4, is characterized in that: described base plate is polystyrene component or tygon component.
9. omnidistance C reactive protein immunochromatographiassay assay quantitative detection test paper according to claim 4, is characterized in that: described in get stuck and comprise plastics upper casing and plastics lower casing, described plastics upper casing fastens to be formed after on plastics lower casing and gets stuck.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107490692A (en) * | 2016-06-09 | 2017-12-19 | 常州博闻迪医药科技有限公司 | A kind of fluorescence immune chromatography method for quantitatively detecting hs-CRP and lipoprotein phospholipase A2 |
| CN108333374A (en) * | 2018-04-08 | 2018-07-27 | 广州天宝颂原生物科技开发有限公司 | C reactive protein, serum amyloid A protein immunochromatography quantify combined detection test paper and preparation method thereof |
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- 2014-10-28 CN CN201420645294.9U patent/CN204287198U/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107490692A (en) * | 2016-06-09 | 2017-12-19 | 常州博闻迪医药科技有限公司 | A kind of fluorescence immune chromatography method for quantitatively detecting hs-CRP and lipoprotein phospholipase A2 |
| CN108333374A (en) * | 2018-04-08 | 2018-07-27 | 广州天宝颂原生物科技开发有限公司 | C reactive protein, serum amyloid A protein immunochromatography quantify combined detection test paper and preparation method thereof |
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