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d-dimer detection kit and detection method

A technology for detecting kits and dimers, which is applied in measuring devices, instruments, and disease diagnosis. It can solve the problems of unsatisfactory rapid detection, poor quantitative accuracy, and long operation time, and achieve clear clinical guiding significance, rapid response, and The effect of high sensitivity

Active Publication Date: 2016-09-28
WEIHAI NEOPROBIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Latex agglutination method is simple and fast, and is suitable for point-of-care testing (POCT), but it can only be used for qualitative determination, mostly for screening; ELISA method has high sensitivity, but poor quantitative accuracy, long operation time, and automatic process Low, not suitable for the needs of POCT; latex particle enhanced immunoturbidimetry (PETIA) can be easily applied to automatic biochemical analyzers, but requires instruments, takes a long time, and is suitable for processing a large number of samples. For emergency departments and primary hospitals, It cannot meet the purpose of rapid detection; the immunogold standard method is similar to the latex agglutination method, which has the advantages of simple and fast operation, but also has poor quantitative accuracy, especially poor repeatability, which limits its clinical application, especially not suitable for Quantitative detection of body fluid marker proteins that need accurate quantification to help diagnose diseases

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Each component of the test paper card in the D-dimer detection kit can be prepared by the following measures:

[0032] 1. Preparation of sample pad 2:

[0033]Soak the glass fiber membrane in the treatment solution containing 1.0% Triton X-100, 2.5% BSA, 0.15M Tris buffer, pH7.5, soak at 4°C for 4 hours, then place it in an oven, and dry it at 37°C 2 hours.

[0034] 2. Preparation of binding pad 3 for absorbing fluorescent microsphere-labeled antibody:

[0035] Soak the glass fiber membrane in 150mM Tris-HCL treatment solution (containing 1.0% Triton X-100, 2.5% BSA, pH7.4), soak at 4°C for 2 hours, then take it out of the oven at 37°C and dry it for 4 hours, and set it aside. Put the glass fiber membrane on the Bio-DotXYZ3050 three-dimensional spraying platform, use the Bio-Jet Quanti300 non-contact micro-quantitative nozzle to spray the D-dimer monoclonal antibody labeled with rare earth fluorescent microspheres on the glass fiber membrane, and dry at 37°C for 1 hou...

Embodiment 2

[0044] Embodiment 2: accuracy test

[0045] Select the above test paper card and fluorescence immunochromatography analyzer (model: NEO-007),

[0046] Setting of the parameters of the fluorescence immunoassay analyzer: after setting the process parameters of the test paper card on the fluorescence immunoassay The D dimer calibrator is measured with a test paper card to obtain the fluorescence intensity value of each calibrator, and the result is input into the parameters of the analyzer to complete the setting of the parameters of the analyzer.

[0047] Main testing materials: clinical samples obtained from relevant hospitals, a total of 200 latex-enhanced immunoturbidimetric value samples, including 100 serum samples, 100 whole blood samples, and the distribution range of D dimer content is between 0-10mg / L .

[0048] Detection method:

[0049] Step 1: Equilibrate the detection reagent and sample to room temperature, take out the test paper card, and lay it flat;

[0050]...

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Abstract

The invention relates to the technical field of fluorescence immunochromatography in medical immunology and particularly relates to a D-dimer detection kit and a D-dimer detection method. The D-dimer detection kit is provided with a test paper card and is characterized in that the test paper card is sequentially provided with a PVC plate, a sample mat, a combination mat, a nitrocellulose membrane and a water absorption mat from bottom to top, wherein a D-dimer monoclonal antibody marked with rare-earth fluorescent micro-spheres is adsorbed on the combination mat, the diameter of each rare-earth fluorescent micro-sphere is 60nm to 120nm, the rare-earth fluorescent microspheres are doped with a rare-earth lanthanide element, the rare-earth fluorescent microspheres are stable at a ground state, and the fluorescence with wavelength range of 540nm to 600nm can be transmitted under the effect of a 340nm-380nm excitation light source; the monoclonal antibody is a monoclonal antibody mixed after purification and is sourced from a monoclonal antibody cell line aiming at 2 to 6 different D-dimer antigen epitopes. The D-dimer detection kit has advantages of simplicity in operation, rapid reaction, high sensitivity, high specificity and the like.

Description

technical field [0001] The invention relates to the technical field of fluorescence immunochromatography in medical immunology, in particular to a D-dimer detection kit and a detection method capable of rapidly and accurately performing quantitative analysis on D-dimer. Background technique [0002] D-dimer (D-Dimer) is derived from the cross-linked fibrin clot dissolved by plasmin. It is a specific degradation product produced by fibrin monomer cross-linked by activated factor XIIIa and then hydrolyzed by plasmin. Is a specific marker of fibrinolytic process. [0003] Under physiological conditions, the body maintains a dynamic balance of coagulation and fibrinolysis to ensure the timely formation and removal of fibrin, while the normal level of D-dimer in the human body is generally below 200 μg / l. If the coagulation and fibrinolysis systems are activated in vivo, The level of D dimer will increase, so D dimer can be used as one of the molecular markers of hypercoagulable...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/533C07K16/18
CPCG01N33/558G01N2800/226G01N2800/52
Inventor 王有志王鹏浩蔡荣
Owner WEIHAI NEOPROBIO
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