CN205015347U - Myeloperoxidase , 2 quantitative joint inspection test paper strips of lipoprotein phospholipase A - Google Patents
Myeloperoxidase , 2 quantitative joint inspection test paper strips of lipoprotein phospholipase A Download PDFInfo
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- CN205015347U CN205015347U CN201520586291.7U CN201520586291U CN205015347U CN 205015347 U CN205015347 U CN 205015347U CN 201520586291 U CN201520586291 U CN 201520586291U CN 205015347 U CN205015347 U CN 205015347U
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- myeloperoxidase
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Abstract
The utility model discloses a myeloperoxidase, 2 quantitative joint inspection test paper strips of lipoprotein phospholipase A, combine pad, nitrocellulose membranes and the pad that absorbs water including bottom plate, sample pad, fluorescent label thing, the test paper strip is combined pad, nitrocellulose membranes, is absorbed water by sample pad, fluorescent label thing and fills up in proper order the overlap joint and paste and constitute on the bottom plate. The fluorescent label thing combines the pad to contain the myeloperoxidase monoclonal antibody of streptavidin mark fluorescin and biotin mark, the lipoprotein phospholipase A 2 monoclonal antibody of biotin mark, the cellulose nitrate is epimembranal to have first detection line, second detection line and matter accuse line. The utility model discloses not only shortened check -out time greatly, improved detectivity, jointly detected simultaneously and still improved the convenience greatly.
Description
Technical field
The utility model belongs to field of immunodetection, is specifically related to a kind of high sensitivity myeloperoxidase, the quantitative joint inspection test strips of lipoprotein phospholipase A2.
Background technology
Myeloperoxidase (Myeloperoxidase, MPO) peroxidase is also called, being a kind of important iron content lysosome, being present in the azurophilic granule of myeloid cell (mainly neutrophil leucocyte and monocyte), is myelocytic specificity marker.In first time in 1967, people recognize that MPO is the constituent to the reaction of exotic invasive generation inherent immunity.MPO is a kind of hemochrome albumen, and molecular weight is 118kD, and containing a pair heavy chain and light chain, it synthesized in marrow before granulocyte enters circulation and is present in azurophilic granule, and environmental stimuli can cause neutrophil accumulation, release MPO.MPO participates in the generation of various diseases, as: inflammation, multiple sclerosis, vasculitis and atherosclerotic etc.Under given conditions, MPO catalytic reaction generates excessive oxygenant, when exceeding the defense reaction of topical antioxidant, oxidative stress and oxidation tissue damage will be caused, MPO regulates inflammatory reaction by number of ways, the most important thing is by regulating nitrogen monoxide to the effect of blood vessel signal transmission and diastole, the directly inflammatory reaction of change blood vessel.
Coronary heart disease (coronaryheartdisease, CHD) is that coronary artery generation is atherosis, spasm, plaque rupture and thrombosis occurs and causes tube chamber to block, cause the disease of coronary insufficiency, myocardial ischemia or infarct.Research finds, developing into unstable angina (UAP) from stable angina pectoris (SAP), develop into acute myocardial infarction AMI (AMI) from UAP is again consecutive variations pathologic process from light to heavy, CHD different pathological stages, its atherosclerotic plaque has different pathological characters.Research confirms, inflammatory reaction all has vital role in atherosclerotic all stages, is formed into vulnerable plaque breaks from fatty streaks.Inflammatory cell plays ascendancy in Earlier atherosclerotic lesion, and its effector molecule accelerates lesion growth, and the activation that causes inflammation causes vulnerable plaque to break, and causes ACS.The display of increasing Clinical Evidence, coronary heart disease pathology affair and inflammation mark change and are closely related, and marker of inflammation is in diagnosis of coronary heart disease, risk stratification, treatment and all have important value in predicting.MPO is leukocytic function and activation marker, is the hemoprotein enzyme containing blood red mutual prothetic group of neutrophil leucocyte, macrophages secrete.Clinical and epidemiological study finds, the atherosclerotic pathologic process of MPO wide participation.Zhang etc. find that patients with coronary heart disease MPO level is higher than control group the earliest.Wainstein etc. further research show that MPO and the CHD order of severity is correlated with, and infiltrate and inflammatory factor expression at a large amount of neutrophil leucocyte and generation phagocyte at Vulnerable plaque.
Lipoprotein phospholipase A2 (Lipoprotein-associatedphospholipaseA2, Lp-PLA2) be phospholipase A 2 (phospholipaseA2, PLA2) a member in superfamily, a kind of serine lipase be made up of 441 amino acid residues, relative molecular mass is 45.4kD.PLA2 is the enzyme race of the short chain acyl on a class energy hydrolytic phosphatide glycerine support Sn-2 position.According to the position existed, substrate specificity, the demand of accessory factor and physiological role etc., PLA2 is mainly divided into four classes: 1. secreting type; 2. endochylema type; 3. non-calcium ionic dependent type; 4. other: take Lp-PLA2 as representative.Lp-PLA2 is primarily of the macrophage of maturation and lymphocyte synthesis and secretion, and be subject to the adjustment of inflammatory mediator, Lp-PLA2 in people's circulation exists with the form be combined with hdl particle, wherein 2/3 be combined with low-density lipoprotein (LDL), 1/3 combines with high-density lipoprotein (HDL) (HDL), very low density lipoprotein (VLDL) VLDL (VLDL).
Lp-PLA2, as a kind of new inflammatory reaction mark, all plays effect in several Main Stage of atherosclerotic (atherosclerosis, AS).Lp-PLA2 produces primarily of macrophage and lymphocyte, in early days about in the research of Lp-PLA2 and As because Lp-PLA2 can be hydrolyzed pro-inflammatory cytokine as platelet activating factor (plateletactivatingfactor, and the relevant oxidized phospholipids of structure PAF), therefore be once considered to suppress inflammatory reaction, even can suppress atherosclerotic formation, therefore be also referred to as platelet-activating factor acetylhydrolase (plateletactivatingfactoracetylhydrolase, PAF-AH).But research has in recent years confirmed that effect that Lp-PLA2 is larger is to promote generation and the development of AS.
In sum, MPO, Lp-PLA2 are inflammatory reaction mark, all take part in coronary atherosclerosis effect, both diagnostic values are incomplete same, joint-detection can be differentiated preferably and diagnose CHD different pathological stages, contribute to the risk stratification of CHD, can also more promptly and accurately Diagnosing Cardiac situation, for the auxiliary diagnosis of angiocardiopathy.
Temporarily without like product on domestic market.
Biotin-avidin system (biotinavidinsystem, 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, and (binding constant is up to 10 to have very high affinity to biology
15m
-1).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 myeloperoxidase, the quantitative joint inspection fast diagnosis reagent of lipoprotein phospholipase A2 and there is no report.
Utility model content
The purpose of this utility model, be to provide a kind of myeloperoxidase, lipoprotein phospholipase A2 quantitative joint inspection test strips, it is based on double antibody sandwich method, fluorescence immunoassay lateral chromatography and biotin-Streptavidin amplification system, provides high sensitivity myeloperoxidase, lipoprotein phospholipase A2 quantitative joint inspection test strips.When using the utility model, improve detection sensitivity and detect stability, reduce non-specific binding, joint-detection increases substantially accuracy and the convenience of detection, is not more than 10 minutes detection time, substantially increases diagnosis efficiency.
The solution that the utility model solves its technical matters is: myeloperoxidase, the quantitative joint inspection test strips of lipoprotein phospholipase A2, 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 biotin labeled myeloperoxidase monoclonal antibody, the fluorescin of biotin labeled lipoprotein phospholipase A2 monoclonal antibody and marked by streptavidin, nitrocellulose filter in described detection zone is fixed with the first detection line identifying the monoclonal antibody of myeloperoxidase another one epi-position and form, the nature controlling line identifying the second detection line that the monoclonal antibody of lipoprotein phospholipase A2 another one epi-position is formed and sheep anti-mouse igg polyclonal antibody formation.
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) utilize the utility model to carry out myeloperoxidase, the detection time of lipoprotein phospholipase A2 is not more than 10 minutes, it is 50.0-8000.0pmol/L that myeloperoxidase detects the range of linearity, it is 20.0-1000.0ng/mL that lipoprotein phospholipase A2 detects the range of linearity, 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) the utility model gets stuck and 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.
(9) the utility model adopts joint-detection, and a detector bar detects two simultaneously, detects convenient.
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, myeloperoxidase, the quantitative joint inspection test strips of lipoprotein phospholipase A2, 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, described fluorescent marker pad 2 is fixed with biotin labeled myeloperoxidase monoclonal antibody (concentration 0.3 ~ 1.5mg/mL), the fluorescin of biotin labeled lipoprotein phospholipase A2 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 ~ 2.0mg/mL), nitrocellulose filter 3 in described detection zone is fixed with the first detection line T1 (concentration 0.5 ~ 3mg/mL) identifying the monoclonal antibody of myeloperoxidase another one epi-position and form, the nature controlling line C (concentration 0.2 ~ 2.0mg/mL) identifying the second detection line T2 (concentration 0.5 ~ 3mg/mL) that the monoclonal antibody of lipoprotein phospholipase A2 another one epi-position is formed and sheep anti-mouse igg polyclonal antibody formation, nature controlling line C is used for the validity of test strip.
The fluorescin that biotin labeled myeloperoxidase monoclonal antibody, biotin labeled lipoprotein phospholipase A2 monoclonal antibody and Streptavidin (or Avidin) mark is fixed on fluorescent marker pad 2, will the monoclonal antibody of identification myeloperoxidase another one epi-position is had, identify that the monoclonal antibody of lipoprotein phospholipase A2 another one epi-position and sheep anti-mouse igg polyclonal antibody are fixed on nitrocellulose filter as detection line T1, T2 and nature controlling line C.When testing sample is added to after in sample pad 1, moved forward by chromatography effect, myeloperoxidase in sample, lipoprotein phospholipase A2 distinguishes myeloperoxidase enzyme antibody (Mab-MPO*Fluoro) that is corresponding and combined with fluorescent label (fluorescin) on fluorescent marker pad 2, lipoprotein phospholipase A2 antibody (Mab-Lp-PLA2*Fluoro) reaction of combined with fluorescent label (fluorescin) forms compound MPO-Mab-MPO*Fluoro, Lp-PLA2-Mab-Lp-PLA2*Fluoro, under chromatography effect, react compound continue to be advanced past MPO antibody nitrocellulose membrane wrapping quilt, time Lp-PLA2 antibody (detection line), the MPO antibody that reaction compound is corresponding coated respectively, Lp-PLA2 antibody capture forms compound (Mab-MPO-MPO-Mab-MPO*Fluoro, Mab-Lp-PLA2-Lp-PLA2-Mab-Lp-PLA2*Fluoro) (detection line), 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 myeloperoxidase in sample, the concentration of lipoprotein phospholipase A2, obtain myeloperoxidase, lipoprotein phospholipase A2 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 2.5 ~ 5.0mm.
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 60 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. myeloperoxidase, the quantitative joint inspection test strips of lipoprotein phospholipase A2, 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 biotin labeled myeloperoxidase monoclonal antibody, the fluorescin of biotin labeled lipoprotein phospholipase A2 monoclonal antibody and marked by streptavidin, nitrocellulose filter in described detection zone is fixed with the first detection line identifying the monoclonal antibody of myeloperoxidase another one epi-position and form, the nature controlling line identifying the second detection line that the monoclonal antibody of lipoprotein phospholipase A2 another one epi-position is formed and sheep anti-mouse igg polyclonal antibody formation.
2. myeloperoxidase according to claim 1, the quantitative joint inspection test strips of lipoprotein phospholipase A2, 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. myeloperoxidase according to claim 2, the quantitative joint inspection test strips of lipoprotein phospholipase A2, 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 quantitative joint inspection test strips of the myeloperoxidase according to any one of claims 1 to 3, lipoprotein phospholipase A2, 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. myeloperoxidase according to claim 4, the quantitative joint inspection test strips of lipoprotein phospholipase A2, is characterized in that: described view window is slot.
6. myeloperoxidase according to claim 4, the quantitative joint inspection test strips of lipoprotein phospholipase A2, is characterized in that: described fluorescin can be the one in green fluorescent protein, phycobniliprotein etc.
7. myeloperoxidase according to claim 4, the quantitative joint inspection test strips of lipoprotein phospholipase A2, is characterized in that: described sample pad is glass fibre component dry after surfactant damping fluid immersion treatment.
8. myeloperoxidase according to claim 4, the quantitative joint inspection test strips of lipoprotein phospholipase A2, is characterized in that: described base plate is polystyrene component or tygon component.
9. myeloperoxidase according to claim 4, the quantitative joint inspection test strips of lipoprotein phospholipase A2, 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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| CN201520586291.7U CN205015347U (en) | 2015-07-30 | 2015-07-30 | Myeloperoxidase , 2 quantitative joint inspection test paper strips of lipoprotein phospholipase A |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108273575A (en) * | 2018-02-26 | 2018-07-13 | 北京华科泰生物技术有限公司 | It is a kind of to be used to diagnose joint-detection micro-fluidic chip of anemic disorders and its preparation method and application |
| 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 |
| CN108398559A (en) * | 2018-02-26 | 2018-08-14 | 北京华科泰生物技术有限公司 | A kind of joint-detection micro-fluidic chip and its preparation method and application for diagnosis of coronary heart disease |
-
2015
- 2015-07-30 CN CN201520586291.7U patent/CN205015347U/en active Active
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108273575A (en) * | 2018-02-26 | 2018-07-13 | 北京华科泰生物技术有限公司 | It is a kind of to be used to diagnose joint-detection micro-fluidic chip of anemic disorders and its preparation method and application |
| CN108398559A (en) * | 2018-02-26 | 2018-08-14 | 北京华科泰生物技术有限公司 | A kind of joint-detection micro-fluidic chip and its preparation method and application for diagnosis of coronary heart disease |
| CN108273575B (en) * | 2018-02-26 | 2020-05-22 | 北京华科泰生物技术股份有限公司 | Combined detection microfluidic chip for diagnosing anemia diseases and preparation method and application thereof |
| 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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