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Fluorescence immunochromatographic detection kit for measuring ST2 and preparation method thereof

A fluorescence immunochromatography, detection kit technology, applied in biological testing, measuring devices, analytical materials, etc., can solve problems such as lack of instruments, and achieve the effects of accurate measurement, fast results, and easy operation

Inactive Publication Date: 2018-12-04
WEIHAI NEOPROBIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the shortcomings and deficiencies of the existing detection products, the present invention proposes a method that combines the advantages of rapid qualitative determination with colored microspheres and accurate quantification with fluorescent microspheres. The requirements of special inspection scenarios such as instruments and no power supply, and can be combined with a fluorescent immunochromatography analyzer to achieve a highly sensitive, accurate and quantitative determination of ST2 indicators. Fluorescence immunochromatography detection kit for ST2 and its preparation method

Method used

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  • Fluorescence immunochromatographic detection kit for measuring ST2 and preparation method thereof
  • Fluorescence immunochromatographic detection kit for measuring ST2 and preparation method thereof
  • Fluorescence immunochromatographic detection kit for measuring ST2 and preparation method thereof

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Embodiment 1

[0026] The various components of the kit in the immunochromatographic detection kit for qualitative and quantitative determination of ST2 can be prepared by the following measures:

[0027] 1. Preparation of sample pad 2:

[0028] Soak the glass fiber membrane in the treatment solution containing 2.0% Triton X-100, 2% BSA, 0.2M 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.

[0029] 2. Preparation of reagent pad 3:

[0030]Soak the glass fiber membrane in 150mM Tris-HCL treatment solution containing 2.0% Triton X-100, 3.0% BSA, pH7.5, soak at 4°C for 3 hours, then take it out of the oven at 37°C and dry it for 4 hours for later use. 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 mouse anti-human ST2 monoclonal antibody-labeled colored fluorescent microspheres on the glass fiber membrane, and dry at 37°C for 2...

Embodiment 2

[0040] Embodiment 2: color microsphere qualitative test and fluorescent immunoassay accuracy test

[0041] Select the above kit and fluorescence immunochromatography analyzer (model: NEO-007),

[0042] Setting of the parameters of the fluorescence immunoassay analyzer: After setting the process parameters of the kit on the fluorescence immunoassayer, take the above assembled kit and use 2.5, 5, 10, 25, 100, 200, 400ng / ml respectively The ST2 calibrator is measured with a kit to obtain the fluorescence intensity value of each calibrator, and the result is input into the parameters of the analyzer to complete the final setting of the parameters of the analyzer.

[0043] Main testing materials: clinical samples were obtained from relevant hospitals, a total of 200 samples of fixed value (the test used the Presage ST2 enzyme-linked immunosorbent assay kit from CriticalDiagnostics), including 100 serum samples, 100 whole blood samples, ST2 content distribution The interval is betw...

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Abstract

The invention relates to the field of immunochromatography in medical immunology, and in particular to a fluorescent immunochromatographic detection kit for determining ST2. The fluorescent immunochromatographic detection kit is composed of an immunochromatographic test strip and a plastic outer shell, and the immunochromatographic test strip is a test strip composed of a bottom plate, a sample pad, a reagent pad, a nitrocellulose membrane and an absorbent pad. the sample pad, the reagent pad, the nitrocellulose membrane and the absorbent pad are sequentially staggered and fixed on the bottomplate, the reagent pad is coated with colored fluorescent microspheres labeled with a mouse anti-human ST2 monoclonal antibody, a detection line on the nitrocellulose membrane is coated with a rabbitanti-human ST2 polyclonal antibody, a control line is coated with a goat anti-mouse polyclonal antibody, the kit has a qualitative determination boundary concentration of 40-55 ng / ml, a fluorescence immunoassay analyzer has a detection sensitivity of 1 ng / ml and a linear range of 1 to 400 ng / ml. The color fluorescent microspheres can quickly obtain qualitative results and obtain quantitative results later. The kit has the characteristics of simple operation, quick response and high sensitivity, and is helpful for satisfying the requirement of different examination scenes.

Description

technical field [0001] The invention relates to the technical field of fluorescent immunochromatography in medical immunology, in particular to a fluorescent immunochromatographic detection kit for measuring ST2 capable of quickly and accurately performing simultaneous qualitative and quantitative analysis on ST2 and a preparation method thereof. Background technique [0002] ST2 is a member of the IL-1 receptor superfamily. It was first discovered by Tominaga in 1989. It has long been considered an orphan receptor associated with inflammation and immune diseases. In 2005, its specific ligand IL- 33, so the study of ST2 was extended to a new field. The human ST2 gene is located on chromosome 2q12, about 40KD, expressed in mast cells, macrophages, activated helper T cells 2 (Th2), cardiomyocytes and cardiac fibroblasts. There are 4 transcripts of ST2 gene, among which the 2 most important isoforms are transmodel ST2 (ST2L) and soluble ST2 (sST2), which are formed by alternat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533G01N33/58G01N33/558
CPCG01N33/533G01N33/558G01N33/582G01N33/585G01N2800/324G01N2800/325
Inventor 王有志刘衍亮陈萍萍宋璐琳王文亮李红江
Owner WEIHAI NEOPROBIO
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