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Fluorescent encoding magnetic bead-based detection method and detection kit

A detection kit and fluorescent coding technology are used in the application of biological nano-magnetic beads and medical testing, which can solve the problems of complex detection methods, high cost, inability to achieve high-throughput, multi-component automatic detection, etc. The effect of mild magnetic adsorption, automation and detection efficiency

Pending Publication Date: 2019-09-20
SUZHOU BAIYUAN GENT CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Therefore, the technical problem to be solved by the present invention is that the prior art cannot realize high-throughput, multi-component, automatic detection, the detection method is complicated, the cost is high and there are more disadvantages, and a detection method based on fluorescently encoded magnetic beads is provided

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] This embodiment provides a detection kit, including 1) fluorescently encoded microspheres coated with mouse anti-human alpha-fetoprotein (AFP) monoclonal antibody; 2) quantum dot-labeled detection antibody; 3) positive control substance; 4 ) Negative control substance; 5) Phosphate buffer solution of pH7.2-7.4.

[0035] Wherein, the fluorescent coded microspheres are embedded with a mass fraction of 2% Fe 3 o 4 Superparamagnetic nanoparticles, polystyrene microspheres with a particle size of 15-20um, coated with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-Hydroxysulfosuccinimide (NHS) can form a covalent bond with the amino group on the mouse anti-human alpha-fetoprotein monoclonal antibody, and the fluorescent signal molecules for fluorescent coding on the fluorescent coding microspheres In order to adopt amino-modified CdSe / ZnS quantum dots with central emission wavelengths of 560nm and 630nm, the mass ratio of the two central emission wa...

Embodiment 2

[0046] The detection kit and detection method described in this example are basically the same as in Example 1, the only difference is that the positive control substance, the negative control substance and the fluorescently encoded microspheres in the buffer are detected by flow cytometry -Target molecule-detect the fluorescence intensity of the antibody complex, and calculate the content of the molecule to be tested in the sample.

Embodiment 3

[0048] This embodiment provides a detection kit, including, 1) fluorescently encoded microspheres coated with antibodies to 5 different target molecules; 2) 5 corresponding detection antibodies labeled with quantum dots; 3) 5 positive controls 4) Negative control substance; 5) Phosphate buffer solution of pH7.2-7.4;

[0049] The fluorescent coded microspheres are embedded with a mass fraction of 3% Fe 3 o 4 Superparamagnetic nanoparticles, polystyrene microspheres with a particle size of 15-20um, coated with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-Hydroxysulfosuccinimide (NHS) can combine with the amino group on the monoclonal antibody of the target molecule to form a covalent bond. The information on the fluorescently encoded microspheres of the antibodies coated with five different target molecules is as follows:

[0050] Fluorescence-encoded microspheres ①: The fluorescent signal group used for fluorescence encoding is CdSe / ZnS quantum dots...

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Abstract

The invention belongs to the technical field of biological nano magnetic bead application and medical examination and relates to a fluorescent encoding magnetic bead-based detection method. The method comprises a capture step, a compound formation step, a separation step and an excitation and determination step. According to the capture step, the compound formation step and the separation step, fluorescent encoding microspheres are specifically bound with required target molecules; a magnetic column is adopted as a transport tool for a compound so as to transfer the compound to different reaction environments; automatic control can be realized; and rapid detection is realized. According to the excitation and determination step, a fluorescence spectrophotometric detection method, a flow cell detection method and a liquid phase chip detection method, are adopted to realize the simultaneous detection and high-throughput detection of different target molecules, in particular, the flow cell detection method can realize intelligent automatic detection through storing a fluorescence encoding microsphere spectral waveform database.

Description

technical field [0001] The invention belongs to the technical field of bio-nano magnetic beads application and medical inspection, and in particular relates to a detection method and a detection kit based on fluorescence-encoded magnetic beads. Background technique [0002] The objects of clinical medical examination mainly include two categories: cells and small molecular substances. Small molecular substances include proteins or nucleic acids. Common detection instruments for cell samples include flow cytometers, hematology analyzers, etc., while detection instruments for small molecule substances include chemiluminescence detectors, chromatography analyzers, etc. In practical applications, the detection of small molecular substances, such as tumor markers, is commonly used for single-item detection by chemiluminescence. However, in a clinical sense, tumor markers and tumors are not completely corresponding. A correlation, the detection of a single tumor marker, the tumor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/52G01N33/58G01N15/14
CPCG01N33/5434G01N33/52G01N33/582G01N33/587G01N15/1434
Inventor 车团结杨涛汪照炎蒋刈李春吴玲贾欢
Owner SUZHOU BAIYUAN GENT CO LTD
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