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Two-in-one determination kit for serum amyloid protein A and C-reactive proteins and preparation method

A technology for amyloid and reactive protein, applied in the field of two-in-one assay kits for serum amyloid A and C-reactive protein, which can solve the problems of high intensity and strong interference, and achieve rapid response, high sensitivity, and clear clinical The effect of instruction

Pending Publication Date: 2017-07-18
WEIHAI NEOPROBIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the usual fluorescence measurement, because the test sample contains a variety of fluorescent components, the background fluorescence (scattered light caused by colloidal particles and solvent molecules in the sample and non-specific fluorescence emitted by proteins and other compounds in the serum) has a large intensity and interference. Strong, become the bottleneck of large-scale promotion of fluorescence analysis

Method used

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  • Two-in-one determination kit for serum amyloid protein A and C-reactive proteins and preparation method
  • Two-in-one determination kit for serum amyloid protein A and C-reactive proteins and preparation method
  • Two-in-one determination kit for serum amyloid protein A and C-reactive proteins and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Each component of the test paper card in the serum amyloid A and C-reactive protein two-in-one assay kit can be made by the following measures:

[0042] 1. Preparation of sample pad 2:

[0043] 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. 2. Preparation of binding pad 3 for absorbing fluorescent microsphere-labeled antibody:

[0044] 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 a 37°C oven and dry it for 4 hours, and set it aside. Put the glass fiber membrane on the Bio-DotXYZ3050 three-dimensional spraying platform, and use the Bio-Jet Quanti300 non-contact micro-quantitative nozzle to mix the two conjugated complexes of serum amyloid A and C-reactive protein labeled with rare earth fluorescent micros...

Embodiment 2

[0053] Embodiment 2: accuracy test

[0054] Select the above test paper card and fluorescence immunochromatography analyzer (model: NEO-007),

[0055] Setting of the parameters of the fluorescence immunoassay analyzer: After setting the process parameters of the test paper card on the fluorescence immunoassay SAA, 0.5, 5, 20, 50, 100, 150, 200mg / L CRP calibrator, use the test paper card to measure, get the fluorescence intensity value of each calibrator, input the result into the parameters of the analyzer, and complete the analyzer parameter settings.

[0056] Main testing materials: clinical samples obtained from relevant hospitals, a total of 300 latex-enhanced immunoturbidimetric value samples, including 100 serum samples, 100 plasma samples, 100 whole blood samples, serum amyloid A and C-reactive protein The content distribution intervals are: SAA: 0.1-40ng / mL, CRP: 0.5-200mg / L.

[0057] Detection method:

[0058] Step 1: Equilibrate the detection reagent and sample t...

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Abstract

The invention discloses a two-in-one determination kit for serum amyloid protein A and C-reactive protein. A test strip is arranged on the determination kit, and the determination kit is characterized in that a PVC board, a sample pad, a combined pad, a nitrocellulose membrane and a water absorbent pad are sequentially arranged on the test strip from bottom to top, wherein two monoclonal antibody-micro-spherical coupling compounds of the anti-serum amyloid protein A and anti-C-reactive proteins which are respectively marked by rare earth Eu<3+> fluorescent microspheres are adsorbed on the combined pad; the diameter of the rare earth fluorescent microspheres is 150nm; the fluorescent microspheres contain rare earth lanthanide elements Eu<3+> and is stable in a basic state, and fluorescence with wavelength of 615nm can be emitted under an action of an excitation light source with the length of 337nm; the monoclonal antibodies are the purified and mixed monoclonal antibodies, which are respectively originated from monoclonal antibody cell strains for 2-6 different serum amyloid protein A and C-reactive protein antigenic epitopes. The two-in-one determination kit has the advantages of convenient operation, quick reaction, high sensitivity, strong specificity and the like.

Description

[0001] Technical field: [0002] The invention relates to the technical field of fluorescent immunochromatography in medical immunology, in particular to a rapid and accurate detection of serum amyloid A and C-reactive protein I in serum, plasma and whole blood samples simultaneously. A two-in-one assay kit for serum amyloid A and C-reactive protein for quantitative analysis of related indicators and a preparation method. [0003] Background technique: [0004] Serum amyloid A (SAA) belongs to the acute phase protein of the human body and exists at a low level in the blood. When the human body is infected by bacteria or viruses, it rises rapidly after inflammation or active lesions or tissue damage. It can rise rapidly during the acute phase of inflammation or infection and decline rapidly during the recovery phase of the disease. Studies have shown that the increase of SAA in serum can be detected in diseases such as bacteria, fungi, virus infection, atherosclerosis, cardiova...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/558G01N21/64
CPCG01N21/6428G01N21/6486G01N33/558G01N33/577G01N33/6893G01N2333/4737G01N2333/47G01N2800/7095
Inventor 王有志王鹏浩李红江
Owner WEIHAI NEOPROBIO
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