WO2013149597A1 - Procédé, système et papier réactif à puce pour une détection parallèle sur divers marqueurs cardiaques - Google Patents

Procédé, système et papier réactif à puce pour une détection parallèle sur divers marqueurs cardiaques Download PDF

Info

Publication number
WO2013149597A1
WO2013149597A1 PCT/CN2013/073796 CN2013073796W WO2013149597A1 WO 2013149597 A1 WO2013149597 A1 WO 2013149597A1 CN 2013073796 W CN2013073796 W CN 2013073796W WO 2013149597 A1 WO2013149597 A1 WO 2013149597A1
Authority
WO
WIPO (PCT)
Prior art keywords
signal
membrane
detection
marker
ligand
Prior art date
Application number
PCT/CN2013/073796
Other languages
English (en)
Chinese (zh)
Inventor
罗朝领
茅柳娟
Original Assignee
安友医疗科技（武汉）有限责任公司
上海领潮生物科技有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 安友医疗科技（武汉）有限责任公司, 上海领潮生物科技有限公司 filed Critical 安友医疗科技（武汉）有限责任公司
Publication of WO2013149597A1 publication Critical patent/WO2013149597A1/fr

Links

Classifications

    • 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
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • 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
    • G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors

Definitions

  • the invention belongs to the technical field of in vitro immunoassay, and relates to a method for parallel detection of markers, in particular to a method for parallel detection of multiple myocardial markers; the invention also relates to a parallel detection system for multiple myocardial markers; It also relates to a chip test strip for parallel detection of multiple myocardial markers. Background technique
  • Cardiovascular diseases such as acute myocardial injury, chronic heart failure and atherosclerosis are common diseases that seriously endanger human health.
  • AMI acute myocardial infarction
  • these enzymes are not unique to the myocardium. They are also abundant in other organs and muscles of the human body. Except for AMI, exercise and inflammation can also be elevated, and the molecular weight of these myocardial enzymes is large, from necrotic tissue into blood. It is slower than some small molecular substances, and the enzyme has a short activity time and a short window time. The value of help for clinical diagnosis is often limited.
  • myocardial marker detection indicators have been more commonly used in clinical laboratory diagnosis, such as troponin T or I, myoglobin, creatine, and drunkenness. (isoenzyme of creatine kinase containing M and B subunits , CK-MB), B-type natriuretic peptide, C-reactive protein (CRP) and the like.
  • the correct application of these new myocardial markers has revolutionized clinical accurate diagnosis, differential diagnosis and judgment of therapeutic effects. Since these new indicators have only been gradually popularized and applied in China in recent years, it is a prerequisite for clinical use and selection of biological characteristics related to them.
  • Myocardial injury markers are mainly troponin T, troponin 1 , creatine kinase isoenzyme mass (CK-MB mass ) and myoglobin ( Myoglobin ).
  • Troponin consists of three subunits, each with its own structure and different regulatory roles. Cardiac troponin regulates the interaction between actin and myosin in the presence of calcium ions, thereby maintaining the myocardium. Relaxation and contraction. When the myocardial cell membrane is intact, cardiac troponin does not penetrate the cell membrane and enters the blood circulation. Only when the myocardial cell membrane is destroyed, troponin can be translated into the blood. The concentration of troponin in the blood circulation is directly proportional to the process of myocardial damage and has a half-life of up to 15 days, which is the best indicator for retrospective testing.
  • CK is an important energy metabolism enzyme in cells. It is widely distributed, and it is the most abundant in muscle cells. It consists of two subunits. CK-MB is mainly present in the outer layer of cardiomyocytes, and has been the clinical diagnosis of myocardial injury. The most specific enzyme in the enzyme language, but there have been many interference factors for measuring enzyme activity by immunosuppression. The sensitivity and specificity of the test are greatly affected. It is recommended by the American Heart Association and the European Heart Association. The chemiluminescence method can determine the quality of CK-MB without being affected by the enzyme activity, and directly detecting the concentration of CK-MB molecules can be more sensitive and specific for clinical help.
  • Myoglobin is mainly found in striated muscle (myocardium, skeletal muscle) cells and plays an important role in the oxidative function of cell membranes. Because it is a small molecule (relative molecular mass 17000 ⁇ 18000), when cardiomyocytes are damaged, Mb is the first marker of organisms that enter the bloodstream, and spreads into the blood faster than CK-MB mass or cTnl/cTnT. . However, because myoglobin is also expressed in skeletal muscle, a large amount of myoglobin can also be released when skeletal muscle is damaged, which does not have myocardial specificity.
  • myoglobin be the best early marker for myocardial damage. Because it is a small molecule, it can enter the blood rapidly during acute myocardial infarction (AMI), so within 1.5 to 6 hours of AMI, Early detection of acute myocardial infarction can be made by dynamically detecting secondary serum myoglobin levels. If the second detection value is significantly higher than the first detection value, it has a very high positive predictive value; if there is no difference between the dynamic detection secondary measurement values, it has a negative predictive value of 100%, excluding the possibility of acute myocardial infarction. Sex.
  • myoglobin may be elevated in severe shock, severe extensive trauma, end-stage renal insufficiency, myocarditis, acute infection, myositis, or myopathy. Therefore, attention should be paid to the differential diagnosis of acute myocardial infarction. Since the window time of myoglobin is the shortest, it is only 3 ⁇ 4d, so this indicator cannot be used for retrospective analysis after the disease occurs.
  • cTnl/cTnT has been consistently recognized by the United States and the European Heart Association as a diagnostic marker for the diagnosis of acute myocardial infarction with high specificity and sensitivity.
  • cTnl/cTnT released from the cytoplasm was rapidly released, and serum/plasma levels increased at 4-6 h.
  • serum/plasma levels increased at 4-6 h.
  • the level of cTn in serum/plasma peaked at 8 to 14 hours after AMI, and decreased to normal after 1 to 2 weeks.
  • cTnl/cTnT has myocardial specificity
  • patients with chest pain 4 hours later can be directly tested with cTnl/cTnT, and its serum/plasma levels are diagnostically specific.
  • Early diagnosis of AMI can be valuable for patients' treatment. time.
  • the detection of cTnl/cTnT is currently the best auxiliary diagnostic indicator.
  • cTnl/cTnT can be used as a monitoring indicator for reperfusion after clinical thrombolysis.
  • cTnl/cTnT is used to determine the accuracy of clinical diagnosis of acute myocardial injury, the late retrospective diagnosis of patients who have not been diagnosed in time, the degree of myocardial injury with skeletal muscle and myocardial injury, and the reperfusion of thrombolytic therapy.
  • Efficacy assessment, assessment of myocardial damage and repair during cardiac surgery are very useful and new diagnostic indicators.
  • the sensitivity and specificity of CK-MB mass detection are much higher than CK-MB activity (activity), so when cTnl or cTnT detection is not performed, CK-MB mass can be used to assist clinical diagnosis for the diagnosis of acute coronary syndrome. And assess the extent of myocardial damage, as well as sensitivity and specificity Close to troponin.
  • B-BNP B-type brain natriuretic peptide
  • B-type natriuretic peptide also known as Brain natriuretic peptide (BNP)
  • BNP Brain natriuretic peptide
  • the BNP secreted by cardiomyocytes first exists as a precursor of 108 amino acids.
  • the cardiomyocytes are stimulated, they are cleaved by the activating enzyme into an inactive linear peptide consisting of 76 amino acids and 32 amino acids.
  • the active cyclic polypeptide, released into the blood circulation, is called NT-proBNP and BNP, respectively.
  • NT-proBNP The biological half-life of NT-proBNP is 60-120 min, while BNP is only 20 min.
  • the translation of B-BNP is closely related to the degree of heart failure, the degree of heart failure is aggravated, and the translation of B-BNP is increased.
  • the main biological role of B-BNP is to participate in sodium regulation, promote urinary sodium excretion and diuresis, dilate blood vessels, maintain a dynamic balance of blood pressure, and antagonize the renin-angiotensin-aldosterone system to increase cardiac output. Published studies have shown that B-BNP levels have an excellent negative correlation with left ventricular ejection fraction, and that B-BNP can be used as an alternative indicator of left ventricular ejection fraction.
  • B-BNP is mainly used to diagnose heart failure, monitor disease progression, evaluate efficacy and prognosis, and is used in patients with AMI after treatment. The recovery of ventricular function was assessed.
  • the BNP level can be significantly decreased, and the BNP level continues to increase or not continuously, which usually indicates that the patient's heart failure has not been corrected or is further aggravated; the differential diagnosis of patients with shortness of breath in the emergency room can also be passed. Determination of B-BNP levels accurately screened for dyspnea caused by non-heart failure patients. Because of its myocardial specificity, B-BNP levels have a high negative predictive value; BNP is used in patients undergoing cardiac surgery. Pre-cardiac function evaluation is also a very important indicator.
  • B-BNP is used for screening high-risk populations, and has important guiding significance, such as diabetes, hereditary heart disease, hypertension, previous myocardial infarction, rheumatic heart disease, and patients undergoing valve replacement surgery. All should be tested regularly for B-BNP to keep abreast of heart function. There are also studies suggesting that elevated B-BNP levels are highly correlated with increased mortality and risk of rehospitalization in high-risk patients, and that B-BNP is considered to be lower than left ventricular ejection fraction (LVEF). And V02 peak has more predictive value. Since B-BNP is currently the only laboratory test for evaluating heart failure, its detection is rapid, sensitive, and specific. The detection of this index to help screen whether or not to perform echocardiography is also a health economic point of view. The best savings option.
  • LVEF left ventricular ejection fraction
  • Coronary heart disease and atherosclerosis are a common type of cardiovascular disease that seriously harms human health and affects the quality of life. As the standard of living increases, its incidence is also growing. Early prevention and intervention of these diseases is an important task for human health.
  • hs-CRP hypersensitive CRP
  • CRP is synthesized by hepatocytes (relative molecular mass 100000 ⁇ 144000), which is normally low in serum/plasma, and CRP content can be multiplied when inflammation or tissue damage is used as the best for inflammation and infection. Laboratory indicators.
  • hs-CRP levels cannot be detected by general immunochemical methods. Ultrasonic latex-enhanced nephelometry can only be used to accurately determine the concentration of hs-CRP in plasma.
  • Foreign researchers have found that hs-CRP levels in healthy people's serum/plasma are less than 0.55 mg/L. When there is a risk of cardiovascular disease, the hs-CRP level is often greater than 2.1 mg/L.
  • hs-CRP is mainly used for screening for risk factors for coronary heart disease.
  • risk factors should not be used as diagnostic indicators.
  • the clinical application value of risk factors should be properly evaluated to effectively help doctors prevent the occurrence and development of coronary heart disease.
  • the technical problem to be solved by the present invention is to provide a method for parallel detection of multiple myocardial markers, which can simultaneously quantitatively detect a plurality of myocardial markers, and can improve the sensitivity of detection and save detection time.
  • the invention also provides a parallel detection system for multiple myocardial markers, which can simultaneously quantitatively detect a plurality of myocardial markers, and can improve the sensitivity of detection and save detection time.
  • the present invention further provides a chip test paper for parallel detection of a plurality of myocardial markers, which can simultaneously quantitatively detect a plurality of myocardial markers, and can improve the sensitivity of detection and save detection time.
  • a method for parallel detection of multiple myocardial markers for detecting myocardial markers part or all of cTNI, cTNT, MYO, CK-MB, BNP, CRP, FABP; the method comprises the following steps:
  • Step S1 fixing the corresponding capture monoclonal antibody of the above-mentioned test substance on the first membrane of the chip test paper, the chip test paper including the first membrane and the second membrane; forming a sandwich detection with the ligands An and Bn
  • the substance to be detected is added from the sample hole, it is moved by diafiltration or siphon or magnetic field or electric field or gravity field to form a first membrane-ligand An-, respectively, in combination with ligands Bn and An.
  • the composite of the analyte-ligand Bn-signal marker is fixed on the first membrane to form a composite array, and is assembled at the same time as the first membrane is captured, leaving a detection hole;
  • Step S2 adding sample serum, when the above-mentioned protein antigen substance is present in the serum of the sample to be added, the labeled secondary antibody on the second membrane of the simultaneous chromatography is combined in the chromatography process, and the combination is firstly Capture antibody capture on the membrane;
  • Step S3 Fixing the fluorescent signal on the first membrane and detecting it by the fluorescence detector, the fluorescence intensity is proportional to the concentration of the target protein in the sample serum, and the fluorescence intensity is converted into a digital signal in the detector, and is directly used according to the internal standard curve. To calculate the protein concentration.
  • the step S3 includes:
  • the laser emitter of the fluorescence detector emits laser light, which simultaneously excites a plurality of signal markers in the chip test paper, and the markers are excited by the laser to generate an energy transition, and the other variable light wave or electronic substance is released;
  • the signal receiving device of the fluorescence detector receives the light wave or electron transit signal decoded by the signal marker, and converts the signal into a digital signal, and the magnitude of the signal marker translation signal is positively correlated with the magnitude; then the fluorescence detector will The processed received signal is calculated, then converted into an analog signal, and the analog signal is output. The output analog signal is compared with the analog calculation, and finally the concentration of the detected substance is output.
  • the signal marker comprises: fluorescein, fluorescent protein, nano gold, silver, nano fluorescent microspheres, quantum dots, rare earth elements, radioactive elements, small molecule chemicals or pigments. kind or more.
  • the fluorescein comprises one or more of FITC, Cy2TM, AlexaTM 488, Cy3TM, AlexaTM 546.
  • the fluorescent protein comprises one or more of GFP, R-PE, and R-PC.
  • a chip test paper for parallel detection of a plurality of myocardial markers for detecting myocardial markers part or all of cTNI, cTNT, MYO, CK-MB, BNP, CRP, FABP;
  • the chip test paper comprises a first film and a second film; a ligand An of each marker is disposed on the first film, and a ligand Bn coupled with a signal label is adsorbed on the second film;
  • the substance to be detected which forms a sandwich detection with the ligands An and Bn is added from the sample hole and then moved by a percolation or siphon or a magnetic field or an electric field or a gravity field to form a first film by combining with the ligands Bn and An, respectively.
  • the complex of the ligand An-substrate-ligand Bn-signal marker is immobilized on the first membrane to form a composite array, which is assembled simultaneously with the capture of the first membrane, leaving the detection well.
  • the signal marker comprises: fluorescein, fluorescent protein, nano gold, silver, nano fluorescent microspheres, quantum dots, rare earth elements, radioactive elements, small molecule chemicals or pigments.
  • a chip test paper comprising a first film and a second film; a ligand An of each marker is disposed on the first film, and a ligand Bn coupled with a signal marker is adsorbed on the second film;
  • the corresponding capture monoclonal antibody of the test substance is immobilized on the first membrane of the chip test paper; the test substance which forms a sandwich detection with the ligands An and Bn is added from the sample hole and then passed through a diafiltration or siphon or a magnetic field or an electric field or gravity Movement under field force combined with ligands Bn and An, respectively, to form a composite of the first membrane-ligand An-detector-ligand Bn-signal marker immobilized on the first membrane to form a composite array, and capture
  • the first membrane is assembled at the same time, leaving the detection hole; the sample serum is added dropwise, and when the above-mentioned protein antigen substance is present in the serum of the sample to be added, the labeled secondary antibody on the second membrane
  • a fluorescence detector for detecting a fluorescent signal immobilized on the first diaphragm.
  • the fluorescence intensity is proportional to the concentration of the target protein in the sample serum.
  • the fluorescence intensity is converted into a digital signal in the detector and directly used according to the internal standard curve. Calculate the protein concentration.
  • the signal marker comprises: fluorescein, fluorescent protein, nano gold, silver, nano fluorescent microspheres, quantum dots, rare earth elements, radioactive elements, small molecule chemicals or pigments. kind or more.
  • the laser emitter of the fluorescence detector emits a laser light, which simultaneously excites a plurality of signal markers in the chip test paper, and the markers are excited by the laser to generate an energy transition, and the image is released.
  • a variated light wave or electronic substance The signal receiving device of the fluorescence detector receives the light wave or electron transit signal decoded by the signal marker, and converts the signal into a digital signal, and the magnitude of the signal marker translation signal is positively correlated with the magnitude; then the fluorescence detector will The processed received signal is calculated, then converted into an analog signal, and the analog signal is output. The output analog signal is compared with the analog calculation, and finally the concentration of the detected substance is output.
  • the invention has the beneficial effects that: the multiple detection methods, the system and the used chip test paper proposed by the invention can quantitatively detect a plurality of myocardial markers at the same time, and can improve the sensitivity of the detection and save the detection time.
  • the system of the invention is convenient to use, convenient to carry, and convenient to use the power source, which may be a dry battery or an ordinary AC power source;
  • the present invention is applicable to various detection systems, and may be an immune reaction, and may be a ligand-receptor-binding reaction such as a nucleic acid reaction;
  • the detection reaction of the method is very fast, from the sample collection to the detection results can be completed in less than half an hour, also called bedside detection;
  • the method can simultaneously detect a plurality of biomarkers
  • the method can accurately and quantitatively detect biomarkers, and can also be used for qualitative detection;
  • the method is also suitable for genetic testing, and all reactions that are suitable for ligand-receptor binding.
  • FIG. 1 is a schematic view showing the composition of a detection system of the present invention
  • FIG. 2 is a schematic structural view of a chip test paper of the present invention
  • Figure 3 is a schematic view of the detector in the detection system of the present invention.
  • Figure 4 is a graphical representation of the test strip for the test strip of the test strip. detailed description
  • the invention discloses a parallel detection method and system for multiple myocardial markers, and a chip test paper. Those skilled in the art can learn from the contents of the paper and appropriately improve the process parameters. Special need All similar substitutions and modifications will be apparent to those skilled in the art and are considered to be included in the invention.
  • the method and the application of the present invention have been described by the preferred embodiments, and it is obvious that the method and application described herein may be modified or appropriately modified and combined without departing from the scope of the present invention. The technique of the present invention is applied.
  • Embodiment 1 Embodiment 1
  • the present invention discloses a multi-cardiac marker parallel detection system, which is a common gold standard marker of myocardial: cTNI, cTNT, MYO, CK-MB, BNP, CRP, FABP;
  • the system includes a fluorescence detector 1, a kit 2.
  • the detector's control system 10 controls the various operating states of the detector.
  • the laser emitter 11 emits a laser which simultaneously excites a plurality of signal markers 21 in the chip 2, which are excited by the laser to undergo an energy transition 21 and then release another variant light or electronic substance 23 .
  • the signal receiving device 12 of the detector 1 receives the light wave or electronic transmission object signal 23 decoded by the signal marker, and converts the signal 23 into a digital signal 13.
  • the size of the signal marker translation signal is positively correlated with the magnitude of the value.
  • the instrument's data processing system then calculates the processed received signal and converts it into an analog signal 15, which is output through the control system, the output of the analog signal is compared to an analog calculation, and finally the concentration of the detected substance is output. correspond.
  • FIG. 2 the structure of the chip test paper for parallel detection of multiple myocardial markers of the present invention is shown in FIG. 2; wherein 5 is a fiber membrane (the fiber membrane is used as the first membrane, and the first membrane may also be silicon). Wafer or silicon wafer or plastic sheet or plastic film or metal sheet), 51 is glass fiber (glass fiber as the second film, the second film can also be glass fiber or silica fiber or metal fiber or organic fiber or high Molecular fiber), 52 is a ligand Bn-signal marker, 53 is a sample well, and 54 is a ligand An.
  • 5 is a fiber membrane (the fiber membrane is used as the first membrane, and the first membrane may also be silicon).
  • 51 is glass fiber (glass fiber as the second film, the second film can also be glass fiber or silica fiber or metal fiber or organic fiber or high Molecular fiber)
  • 52 is a ligand Bn-signal marker
  • 53 is a sample well
  • 54 is
  • the chip test paper is used for detecting myocardial markers: part or all of cTNI, cTNT, MYO, CK-MB, BNP, CRP, FABP; and the detected substance which forms a sandwich detection with the ligands An and Bn from the sample hole After the addition, it is separated by the percolation (or siphon, magnetic field, electric field, gravity field) and combined with the ligands Bn and An to form a fiber membrane-ligand.
  • the composite of An-substrate-ligand Bn-signal marker is immobilized on the fiber membrane to form a composite matrix ⁇ ij, which is assembled simultaneously with the capture fiber membrane to leave a detection well.
  • the detection monoclonal antibodies which are paired with the fiber membranes to form a sandwich detection target substance are respectively labeled with a signal marker, and then fixed to the glass fibers, and assembled together with the capture fiber membrane to leave a detection hole.
  • 2 is a chip test paper, and a fiber membrane is disposed on the chip test paper 2, and a ligand 21 is provided on the fiber membrane, and the ligand 21 is branched, and the ligand An is fixed on the substrate
  • the marker 22 is n
  • the ligand 24 is Bn on the marker n
  • the plurality of detected substances form an affinity bond with the solid phase ligand, and one or more signal markers 23 around the top or the top of the ligand.
  • n 1, 2, 3, 4, 5 composite.
  • the complex (n species) is detected on the above detector to detect the intensity of the signal substance and is converted to the concentration of the corresponding substance, thereby detecting the concentration or strength of the detected biomarker (n species).
  • the purpose of rapid diagnosis is achieved.
  • the aforementioned label 23 is: fluorescein (fluorescein: FITC, Cy2TM, AlexaTM 488 : Cy3TM, AlexaTM 546 conventional fluorescein), fluorescent protein (GFP, R-PE, R-PC), nanogold , silver, nano-fluorescent microspheres, quantum dots, rare earth elements, radioactive elements, small molecule chemicals or pigments.
  • the size of the above-mentioned marker 23 translation signal is positively correlated with the magnitude of the value.
  • the aforementioned light-wave or electronic substance which is excited by the laser after being excited by the laser is different from the light wave or electronic substance of the label itself.
  • the moving force of the biomarker n and the signal marker-ligand Bn complex in the aforementioned substance to be detected on the solid phase support is chromatography, siphoning, diafiltration, electrophoresis, magnetophoresis or gravity.
  • the aforementioned fiber membrane may be a PV membrane, a glass slide, a plastic plate or a gel plate.
  • the aforementioned signal marker 23 is fluorescein (fluorescein: FITC, Cy2TM, AlexaTM 488, Cy3TM, AlexaTM 546 conventional fluorescein), fluorescent protein (GFP, R-PE, R-PC), nanogold (Can be nano gold, nano silver, nano copper, etc.), nano fluorescent microspheres, quantum dots, rare earth elements, radioactive elements, small molecule chemicals or pigments.
  • fast test instrument Small size, easy to carry, can use ordinary power and dry battery power, can detect multi-point fluorescence at the same time.
  • ligand Bn-signal marker ligand Bn can be antibody, antigen, biotin-avidin, ligand, nucleic acid, lectin, etc., can be the same substance or different species
  • ligand Bn can be antibody, antigen, biotin-avidin, ligand, nucleic acid, lectin, etc.
  • labeling signal-ligand Bn-detecting substance n labeling signal-ligand Bn-detecting substance n
  • ligand An may be antibody, antigen, organism An avidin, a ligand, a nucleic acid, a lectin, etc., which may be one or more of the same substance or a different species, is immobilized on a solid support, and simultaneously with a marker signal in motion -
  • the ligand Bn - the analyte n forms an affinity bond, thus forming a complex of the ligand An-detected substance n-ligand Bn-signaling substance on the test paper solid support.
  • a siphon (or electrophoresis, magnetic field, etc.) technique to move the sample on a fibrous membrane (or plastic plate, rubber plate, glass plate, etc.) carrier, during the movement of the biomarker and the labeled ligand 23 placed on the carrier (including : an antibody, an antigen, a nucleic acid, a lectin, etc., binds together and moves together to a ligand 24 immobilized on a carrier (ligand 24 may be immobilized by adsorption, or covalent coupling, etc., and the fixation may be point-like, Line shape, etc.)
  • the detector (multi-function fast chip tester) is a very flexible and versatile detection technology platform.
  • the principle is that proteins (such as antigens, antibodies) for different detection substances are bound to specific solid supports (such as slides, fiber membranes, gels, etc.) in micro-column form, and then with the detected object (the measured object can be It is a combination of antigens, antibodies, small molecules of drugs, nucleic acids or enzymes in body fluids or serum, and the detected substances are also associated with signal markers (which may be fluorescein, quantum dots, gold nanoparticles, silver particles, or fluorescent protein molecules, etc.)
  • the labeled protein binds to form a complex of ligand A-detected substance n-ligand B-labeled signal.
  • the instrument emits a fixed-wavelength laser to illuminate the signal with a plurality of fluorescently labeled micro-integral complexes; the label is excited by the laser and then emits another variable fixed-wavelength photon, which is sequentially
  • the receiver of the instrument captures and converts it into a digital signal; the numerical signal of the corresponding complex is calculated by the data processing software to obtain the concentration of the corresponding complex, and the concentrations of the various biomarkers of the detected object are sequentially calculated.
  • Chip test paper production method
  • biomarker n can be antigen, antibody, drug, chemical small molecule, nucleic acid, peptide
  • the ligand An for different analytes is prepared with a certain concentration of biological buffer (which may be lng-10mg), and then the ligand An is sequentially arranged in a micro-column by a spotting device on the solid support (solid support)
  • the material may be a nitrocellulose membrane, a gel, a slide, a plastic plate, etc., which may be in the shape of a long strip, a square, a center, etc., and is subjected to a certain chemical and physical treatment to keep the ligand An as it should be.
  • Biological activity In addition, there is a quality control point for the internal control point (or internal standard) of the corresponding detection object and whether the sample is normally reacted or not.
  • the ligand Bn and the labeled signal substance are linked together by their corresponding physical and chemical properties by physical, chemical or biological means to stabilize them.
  • Chip test paper can have various shapes according to its fluid method.
  • the basic components include: sample sample hole, ligand Bn marker storage position, ligand An microarray and support, sample flow driving force.
  • Device can be absorbent paper, percolating paper, siphoning tank, capillary or electromagnetic field, etc.).
  • the sample which can be detected by the method of the present invention is not particularly limited and may be any sample containing a biomarker, and representative examples include blood sample, serum sample, urine sample, saliva sample, body fluid sample; various food vegetable washing liquid, soaking Liquid; nucleic acid extract, PCR amplification product, and the like.
  • the instrument will automatically obtain the signal of the signal mark substance on the four original composites, and convert it into a digital signal and then calculate it internally by the data processing software to detect the detected sample.
  • the concentration of n biomarkers are recorded by the data output form (or print output form).
  • the detector includes: a laser 31, a filter group 32, a filter 33, a bidirectional color mirror 38, a mirror 35, a first objective lens 34, a second filter 36, and a second objective lens 37. , light bar 40 and chip test paper 39, and ⁇ (photomultiplier tube) components, AMP (signal amplifier) components, converter A / D (signal data converter) components, CD chips, output printing devices, motherboards and power supplies ( Not shown in the figure).
  • ⁇ photomultiplier tube
  • AMP signal amplifier
  • converter A / D signal data converter
  • the optical components are connected in the instrument box in turn, and the power supply and the control board are distributed, and the operation of each line and component is detected in turn to check whether the requirements are met. After the full compliance, the motherboard control program and power supply are turned on to test the system operation.
  • the detector is debugged according to the following indicators:
  • the instrument can add an electric field and or a magnetic field in the chip test socket, so that the charged or magnetic substance on the chip test paper can be oriented under the corresponding force to achieve the moving force of the detected substance.
  • the chip test material can be NC film or glass piece or plastic piece, etc.
  • the chip dot matrix is 5X6 or other arrangement (according to the specific product), the pitch is 1mm, the dot diameter is lOum-500um, and the total width of the test paper is 3-30mm. Length range: 30-100mm.
  • Appearance includes sample hole, chip illumination window, sealing card, positioning mark, etc.
  • the internal structure of the chip test strip includes: filter paper, glass fiber where the signal marker ligand is located, ligand lattice (including test points, calibration points, and quality control points).
  • the chip tester is fully in line with its advantages of fast, sensitive, quantitative detection, multiple joint inspections, and convenient carrying. It is a future application and its wide range of products, which will promote the development of future clinical testing.
  • Embodiment 2
  • cTNI, cTNT, MYO, BNP, CRP, FABP antibody pairs are available for LINC-BIO and Hytest;
  • BNP, cTNI, cTNT, and CRP antigens are also provided by LINC-BIO;
  • the primary antibodies of the six items were prepared at a concentration of l-5 mg/ml with 50 Mm Tris-HCl PH7.5.
  • Rabbit anti-mouse IgG was prepared at a concentration of 1-5 mg/ml with 50 Mm Tris-HCl pH 7.5.
  • NC membrane Whatman company number: BA85
  • the second antibody of the labeled antibody was prepared into a mixed solution of 5-30 ug/ml in 20 mM PBS pH 7.4 buffer;
  • cTNI antigen concentration For: 0, 0.5, 2.5, 12.5, 25, 50 ng/ml six gradients, each volume 3 ml spare; cTNT antigen concentration: 0, 0.5, 2.5, 12.5, 25, 50 ng/ml Six gradients, MYO antigen concentration is : 6 gradients of 0, 10, 50, 250, 1000, 4000 ng/ml, BNP antigen concentration: 0, 1 , 5, 20, 100, 200 ng/ml six gradients, CRP antigen concentration: 0, 0.01, 0.3 Six gradients of 0.9, 3, 9ug/ml, CK-MB antigen concentration: 0, 2.5, 10, 50, 200, 500 ng/ml six gradients, FABP antigen concentration: 0, 2, 10, 50, 100 , 200 ng / ml six
  • Test experiment take out the six test strips of the produced myocardial markers; take the six gradients of the prepared myocardial markers lOOul, add the test holes of the test paper in turn, and insert the chip test paper into the chip tester (Line xMAP)
  • the magnetic field power is turned on in the platform jack, so that it moves in the magnetic field (that is, the sample first enters the chip test paper to add the sample, passes through the sample filter paper, enters the glass fiber layer, and the six antigens in the sample are respectively labeled with the magnetic quantum fluorescent
  • the antibody binds to form a magnetic quantum fluorescent-antibody 2-antigen complex, and the magnetic quantum dots move under the magnetic field of the instrument.
  • the complex When moving to an antibody spot to which six anti-cardiac markers are immobilized, the complex is captured by the immobilized antibody to form: Magnetic quantum fluorescent-antibody 2-antigen-antibody 1 complex, and the complex cannot continue to move, while other uncaptured substances continue to flow to the top absorbent paper.
  • the chip test strip detector Line The xMAP platform reads the fluorescent signal on the complex and repeats the test 20 times per sample per batch. A signal value corresponding to the fluorescence. In turn, force. The force into a test strip. The sample wells were allowed to react for 5 mins and then placed in a Line xMAP instrument to read the fluorescent signals at four points, and each sample of each batch was repeatedly tested three times to record its corresponding fluorescence signal value.
  • Sample collection respectively, collected 270 samples of the same day in the laboratory of three Sancha hospitals, including 30 positive samples of myocardial markers, 30 normal samples, 60 positive samples of myocardial markers, and more than two positive samples of myocardial markers. example.
  • cTNI, MYO, BNP antibody pairs are available for LINC-BIO and Hytest;
  • BNP, cTNI antigen is also provided by LINC-BIO;
  • the primary antibodies of the four items were prepared at a concentration of l-5 mg/ml with 50 Mm Tris-HCl PH7.5.
  • Rabbit anti-mouse IgG was prepared at a concentration of 1-5 mg/ml with 50 Mm Tris-HCl pH 7.5.
  • 1.3 Prepare nitrocellulose membrane (NC membrane, Whatman company number: BA85) 20mmX5mm size.
  • the TM 2.1 chip spotting system places the prepared four primary antibodies and rabbit anti-mouse IgG on the NC membrane, respectively, at a point of 10-200 nl, arranged in 3 rows, and rabbit anti-mouse IgG in the first row.
  • the primary resistance of the four items is divided into two rows, two points in each row, a point spacing of 3 mm, and a row spacing of 2 mm.
  • the film strips with good samples are placed in a vacuum drying oven and air-dried for storage.
  • the second antibody of the labeled antibody was prepared into a mixed solution of 5-30 ug/ml in 20 mM PBS pH 7.4 buffer;
  • Test experiment take out the six test strips of the produced myocardial markers; take the six gradients of the prepared myocardial markers lOOul, add the test holes of the test paper in turn, and insert the chip test paper into the chip tester (Line xMAP)
  • the magnetic field power is turned on in the platform jack, so that it moves in the magnetic field (that is, the sample first enters the chip test paper to add the sample, passes through the sample filter paper, enters the glass fiber layer, and the six antigens in the sample are respectively labeled with the magnetic quantum fluorescent
  • the antibody binds to form a magnetic quantum fluorescent-antibody 2-antigen complex, and the magnetic quantum dots move under the magnetic field of the instrument.
  • the complex When moving to an antibody spot to which six anti-cardiac markers are immobilized, the complex is captured by the immobilized antibody to form: Magnetic quantum fluorescent-antibody 2-antigen-antibody 1 complex, and the complex cannot continue to move, while other uncaptured substances continue to flow to the top absorbent paper.
  • the chip test strip detector Line The xMAP platform reads the fluorescent signal on the complex and repeats the test for each sample in each batch. The corresponding fluorescent signal values were recorded 20 times.
  • cTNI, CRP, BNP antibody pairs are available for LINC-BIO and Hytest;
  • BNP, CtnI, and CRP antigens are also provided by LINC-BIO;
  • the primary antibodies of the three items were prepared at a concentration of l-5 mg/ml with 50 Mm Tris-HCl PH7.5.
  • Rabbit anti-mouse IgG was prepared at a concentration of l-5 mg/ml with 50 Mm Tris-HCl pH 7.5.
  • 1.3 Prepare nitrocellulose membrane (NC membrane, Whatman company number: BA85) 20mmX5mm size.
  • the second antibody of the labeled antibody was prepared into a mixed solution of 5-30 ug/ml in 20 mM PBS pH 7.4 buffer;
  • cTNI antigen concentration 0, 0.5, 2.5, 12.5 , 25, 50 ng / ml six gradients, each volume of 3 ml spare; CRP antigen concentration: 0, 0.01, 0.3, 0.9, 3, 9 ug / ml six gradients, BNP antigen concentration: 0, 1, 5, 20, 100, 200 ng/ml six gradients; C six gradients, 3 ml each for use. And 3 batches were prepared for each gradient.
  • Test experiment take out the six test strips of the produced myocardial markers; take the six gradients of the prepared myocardial markers lOOul, add the test holes of the test paper in turn, and insert the chip test paper into the chip tester (Line xMAP)
  • the magnetic field power is turned on in the platform jack, so that it moves in the magnetic field (that is, the sample first enters the chip test paper to add the sample, passes through the sample filter paper, enters the glass fiber layer, and the six antigens in the sample are respectively labeled with the magnetic quantum fluorescent
  • the antibody binds to form a magnetic quantum fluorescent-antibody 2-antigen complex, and the magnetic quantum dots move under the magnetic field of the instrument.
  • the complex When moving to an antibody spot to which six anti-cardiac markers are immobilized, the complex is captured by the immobilized antibody to form: Magnetic quantum fluorescent-antibody 2-antigen-antibody 1 complex, and the complex cannot continue to move, while other uncaptured substances continue to flow to the top absorbent paper.
  • the chip test strip detector Line The xMAP platform reads the fluorescent signal on the complex and repeats the test for each sample in each batch. The corresponding fluorescent signal values were recorded 20 times.
  • Sample collection respectively, collected 270 samples of the same day in the laboratory of three Sancha hospitals, including 30 positive samples of myocardial markers, 30 normal samples, 60 positive samples of myocardial markers, and more than two positive samples of myocardial markers. example.
  • cTNI, MYO, BNP, FABP antibody pairs are available for LINC-BIO and Hytest; CK-MB, antibody pairs for Medix
  • BNP, cTNI antigen is also provided by LINC-BIO;
  • the quantum fluorescence is a 50 nm quantum dot with an absorption wavelength of 532 nm and an excitation wavelength of 757 nm, which is purchased from a handsome organism.
  • the primary antibodies of the five items were prepared at a concentration of l-5 mg/ml with 50 Mm Tris-HCl PH7.5.
  • Rabbit anti-mouse IgG was prepared at a concentration of l-5 mg/ml with 50 Mm Tris-HCl pH 7.5.
  • NC membrane Whatman company number: BA85
  • the TM 2.1 chip spotting system places the prepared five primary antibodies and rabbit anti-mouse IgG on the NC membrane, respectively, at a point of 10-200 nl, arranged in 2 rows, and rabbit anti-mouse IgG in the upper left corner. Plus the five items of the primary antibody are divided into two rows, each row of 3 points, the point spacing is 2mm, and the row spacing is 3mm. And the film strips with good samples are placed in a vacuum drying oven and air-dried for storage. 2.2 The secondary antibodies for the five projects are labeled with quantum fluorescence:
  • the secondary antibody to be labeled is placed in a dialysis bag, the dialysis bag is tied, and the dialysis bag to be labeled is placed in the above-prepared quantum fluorescent solution and dialyzed overnight at 4 ° C;
  • the second antibody of the labeled antibody was prepared into a mixed solution of 5-30 ug/ml in 20 mM PBS pH 7.4 buffer;
  • cTNI antigen is: 0, 0.5, 2.5, 12.5, 25, 50 ng/ml six gradients, each volume is 3 ml;
  • MYO antigen concentration is: 0, 10, 50, 250, 1000, 4000 ng/ml six gradients, BNP
  • the antigen concentration was: 0, 1 , 5 , 20, 100, 200 ng / ml six gradients;
  • CK-MB antigen concentration 0, 2.5, 10, 50, 200, 500 ng / ml;
  • FABP antigen concentration 0, 2 , 10, 50, 100, 200 ng / ml six gradients, each volume of 3ml spare. And 3 batches were prepared for each gradient.
  • Test experiment take out the six test strips of the produced myocardial markers; take the six gradients of the prepared myocardial markers lOOul, add the test holes of the test paper in turn, and insert the chip test paper into the chip tester (Line xMAP)
  • the magnetic field power is turned on in the platform jack, so that it moves in the magnetic field (that is, the sample first enters the chip test paper to add the sample, passes through the sample filter paper, enters the glass fiber layer, and the six antigens in the sample are respectively labeled with the magnetic quantum fluorescent
  • the antibody binds to form a magnetic quantum fluorescent-antibody 2-antigen complex, and the magnetic quantum dots move under the magnetic field of the instrument.
  • the complex When moving to an antibody spot to which six anti-cardiac markers are immobilized, the complex is captured by the immobilized antibody to form: Magnetic quantum fluorescent-antibody 2-antigen-antibody 1 complex, and the complex cannot continue to move, while other uncaptured substances continue to flow to the top absorbent paper.
  • the chip test strip detector Line The xMAP platform reads the fluorescent signal on the complex and repeats the test for each sample in each batch. The corresponding fluorescent signal values were recorded 20 times.
  • the multi-cardiac marker parallel detection method and system and the used chip test paper proposed by the invention can simultaneously quantitatively detect a plurality of myocardial markers, and can improve detection sensitivity and save detection time.
  • the description and application of the present invention are intended to be illustrative, and not intended to limit the scope of the invention. Variations and modifications of the embodiments disclosed herein are possible, and various alternative and equivalent components of the embodiments are well known to those of ordinary skill in the art. It is apparent to those skilled in the art that the present invention may be embodied in other forms, configurations, arrangements, ratios, and other components, materials and components without departing from the spirit or essential characteristics of the invention. Other variations and modifications of the embodiments disclosed herein may be made without departing from the scope and spirit of the invention.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biotechnology (AREA)
  • Analytical Chemistry (AREA)
  • Cell Biology (AREA)
  • Pathology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Endocrinology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

L'invention concerne un procédé, un système et des papiers réactifs à puce pour une détection parallèle sur divers marqueurs myocardiques. Le papier réactif à puce est utilisé pour détecter une partie ou l'ensemble des marqueurs myocardiques comprenant cTNI, cTNT, MYO, CK-MB, BNP, CRP et FABP ; le papier réactif à puce comprend une première membrane et une seconde membrane ; un ligand An de chaque marqueur est agencé sur la première membrane et un ligand Bn couplé à un marqueur de signal est absorbé sur la seconde membrane ; et un matériau détecté pris en sandwich par les ligands An et Bn est placé à l'intérieur par un trou d'échantillonnage, puis, sous l'effet de la force d'action de percolation ou d'autres facteurs, le matériau détecté se déplace pour être combiné respectivement aux ligands An et Bn pour former une matrice composite de la première membrane - du ligand An - du matériau détecté - du ligand Bn - du marqueur de signal, qui est fixée sur la première membrane, la matrice composite et une membrane à fibre de capture sont assemblées simultanément, et un trou de détection est réservé. Le procédé, le système et le papier réactif à puce décrits par l'invention peuvent être utilisés pour détecter simultanément et quantitativement les divers marqueurs myocardiques; de plus, la sensibilité de détection peut être améliorée et le temps de détection est économisé.
PCT/CN2013/073796 2012-04-06 2013-04-07 Procédé, système et papier réactif à puce pour une détection parallèle sur divers marqueurs cardiaques WO2013149597A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201210100476.3 2012-04-06
CN2012101004763A CN102636651A (zh) 2012-04-06 2012-04-06 多项心肌标志物并行检测方法及系统、芯片试纸

Publications (1)

Publication Number Publication Date
WO2013149597A1 true WO2013149597A1 (fr) 2013-10-10

Family

ID=46621120

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2013/073796 WO2013149597A1 (fr) 2012-04-06 2013-04-07 Procédé, système et papier réactif à puce pour une détection parallèle sur divers marqueurs cardiaques

Country Status (2)

Country Link
CN (1) CN102636651A (fr)
WO (1) WO2013149597A1 (fr)

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102636651A (zh) * 2012-04-06 2012-08-15 上海领潮生物科技有限公司 多项心肌标志物并行检测方法及系统、芯片试纸
CN104698171B (zh) * 2013-12-04 2017-02-01 内蒙古农业大学 一种基于胶体金与发光量子点二聚体的二级可选灵敏度侧向层析快速检测方法
CN106443004A (zh) * 2016-04-12 2017-02-22 上海奥普生物医药有限公司 一种快速定量同时检测cTnI、H‑FABP的时间分辨荧光免疫层析试剂及制备方法
CN206270348U (zh) * 2016-10-14 2017-06-20 深圳市金准生物医学工程有限公司 一种联合诊断试纸组件
CN107643284B (zh) * 2017-09-01 2018-12-18 北京华科泰生物技术有限公司 用于检测心肌酶系列的磁微粒的微流控化学发光检测系统
CN109709318A (zh) * 2018-12-28 2019-05-03 南京祥中生物科技有限公司 同时检测多种心肌标志物的化学发光微阵列芯片及试剂盒
CN110411951B (zh) * 2019-08-13 2021-11-30 信阳师范学院 一种用于同时检测双心肌标志物的光电化学生物传感器的制备方法
CN113655219B (zh) * 2021-08-20 2023-10-13 中国人民解放军军事科学院军事医学研究院 基于上转发光免疫层析技术的crp和saa的联合定量检测方法

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101271108A (zh) * 2008-01-11 2008-09-24 广州益善生物技术有限公司 心肌梗塞早期诊断液相芯片及其制备方法
WO2009104974A2 (fr) * 2008-02-22 2009-08-27 Christopher Joseph Pemberton Biomarqueurs
CN101661034A (zh) * 2008-08-28 2010-03-03 上海领潮生物科技有限公司 同时进行多种生物标志物并行检测的方法和芯片试纸
CN102062735A (zh) * 2009-11-18 2011-05-18 江苏迈迪基因生物科技有限公司 急性冠状动脉综合症的生物标志物检测法及诊断试剂盒
CN102636651A (zh) * 2012-04-06 2012-08-15 上海领潮生物科技有限公司 多项心肌标志物并行检测方法及系统、芯片试纸

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5902731A (en) * 1998-09-28 1999-05-11 Lifescan, Inc. Diagnostics based on tetrazolium compounds
CN1363839A (zh) * 2001-09-28 2002-08-14 上海生物芯片有限公司 早期心肌梗死诊断生物芯片
CN101881775A (zh) * 2010-06-04 2010-11-10 无锡安抗生物科技有限公司 一种肌钙蛋白i和肌钙蛋白t的双通道检测卡
CN102103143B (zh) * 2011-02-24 2013-09-25 南京基蛋生物科技有限公司 一种双指标检测用胶体金试纸条及其制备方法
CN102323422B (zh) * 2011-05-30 2014-03-12 中国科学院上海微系统与信息技术研究所 半定量同时检测cTnI和Myo的免疫层析试纸条及其制备

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101271108A (zh) * 2008-01-11 2008-09-24 广州益善生物技术有限公司 心肌梗塞早期诊断液相芯片及其制备方法
WO2009104974A2 (fr) * 2008-02-22 2009-08-27 Christopher Joseph Pemberton Biomarqueurs
CN101661034A (zh) * 2008-08-28 2010-03-03 上海领潮生物科技有限公司 同时进行多种生物标志物并行检测的方法和芯片试纸
CN102062735A (zh) * 2009-11-18 2011-05-18 江苏迈迪基因生物科技有限公司 急性冠状动脉综合症的生物标志物检测法及诊断试剂盒
CN102636651A (zh) * 2012-04-06 2012-08-15 上海领潮生物科技有限公司 多项心肌标志物并行检测方法及系统、芯片试纸

Also Published As

Publication number Publication date
CN102636651A (zh) 2012-08-15

Similar Documents

Publication Publication Date Title
WO2013149597A1 (fr) Procédé, système et papier réactif à puce pour une détection parallèle sur divers marqueurs cardiaques
CN106461672B (zh) 用于诊断舒张功能障碍的igfbp7
US5358852A (en) Use of calcium in immunoassay for measurement of C-reactive protein
US5149629A (en) Coulometric assay system
CN106596969B (zh) 一种电致化学发光免疫传感器的制备、产品、检测及应用
CN102087293A (zh) 一种全程定量检测肌钙蛋白i的免疫层析试纸条及其制备方法
CN1781022A (zh) 用于进行现场测试的膜条生物传感器系统
Savonnet et al. Recent advances in cardiac biomarkers detection: From commercial devices to emerging technologies
US20120149128A1 (en) Assays and assay devices
CN101661034A (zh) 同时进行多种生物标志物并行检测的方法和芯片试纸
CN101339196A (zh) 量子点标记免疫层析试纸快速检测膀胱癌方法
Liu et al. An improved portable biosensing system based on enzymatic chemiluminescence and magnetic immunoassay for biological compound detection
CN105891508A (zh) 一种快速定量检测h-fabp的时间分辨荧光免疫层析试剂及制备方法
EP1722232A1 (fr) Dispositifs et procédés de diagnostics ou de prévoir d'un stade précoce des dysfonctionnements cardiaques
Cho et al. Two-dimensional paper chromatography-based fluorescent immunosensor for detecting acute myocardial infarction markers
KR101916608B1 (ko) 생화학-면역 하이브리드 바이오센서 및 이를 포함하는 센서시스템
Zhong et al. Recent advances in electrochemical aptasensors for detecting cardiac biomarkers: A review
Vairaperumal et al. Optical nanobiosensor-based point-of-care testing for cardiovascular disease biomarkers
CN102305866B (zh) 快速诊断急性心肌梗死的检测装置
CN104714031A (zh) 一种心肌梗死快速检测试剂盒
Xue et al. A dry chemistry, ultrasensitive microfluidic fiber material-based immunosensor for electrochemiluminescence point-of-care testing of luteinizing hormone
Ming-Qing et al. Research progress of C-reactive protein analysis
Dong et al. Advances in heart failure monitoring: Biosensors targeting molecular markers in peripheral bio-fluids
CN110568192A (zh) 基于氧化锌纳米线荧光增强的纸基分析装置及方法
CN108037281A (zh) 一种心型脂肪酸结合蛋白免疫检测试剂及其制备和检测方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 13771825

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 2015N DATED 27/02/2015)

122 Ep: pct application non-entry in european phase

Ref document number: 13771825

Country of ref document: EP

Kind code of ref document: A1