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A multiple digital elisa detection method and microfluidic chip

A microfluidic chip and detection method technology, applied in chemical instruments and methods, measurement devices, biological tests, etc., can solve the problems of low detection limit, poor sensitivity and specificity, long time consumption, etc., and achieve low time and labor costs, High sensitivity and specificity, low sample consumption

Active Publication Date: 2021-12-17
深圳市科瑞达生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Single target detection consumes a lot of samples, takes a long time, and the cost of labor and reagents is high, which cannot meet the needs of large-scale screening and diagnosis of clinical diseases. However, the detection throughput of existing multiple detection methods is only a few or a dozen Therefore, if the detection throughput can be greatly improved for parallel multi-channel detection, the detection efficiency can be significantly improved and real-time rapid disease monitoring can be realized. In addition, there are also problems of low detection limit and crosstalk between target molecules when performing multiple detection. problems, resulting in poor sensitivity and specificity of detection

Method used

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  • A multiple digital elisa detection method and microfluidic chip
  • A multiple digital elisa detection method and microfluidic chip
  • A multiple digital elisa detection method and microfluidic chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] The process and test results of the detection of various allergen antibodies based on the microfluidic chip provided in Example 1 are as follows:

[0079] (1) Microfluidic chip assembly

[0080] Using modeling software to create a microfluidic chip model, the above-mentioned microfluidic chip was made of polymethyl methacrylate material, sealed with acrylic glue, and the opening sizes of the cubic column barriers in the microfluidic chip were 500 μm, 450 μm, 400 μm, 350 μm, 300 μm, corresponding to the different diameters in step S1, take the polystyrene microspheres of the above five sizes, and modify them with peanut agglutinin, β-lactoglobulin, apple rMal d4, hazelnut rCor a1 and campanula Grass pollen allergens are used to detect the corresponding IgE antibodies in human serum (samples with target molecules). The BSA is sealed to prevent non-specific adsorption. The polystyrene microspheres are poured into the microfluidic chip, and the reciprocating motor is start...

Embodiment 2

[0084] Example 2 provides detection of multiple tumor markers based on the above-mentioned multiple detection microfluidic chip.

[0085] (1) Microfluidic chip assembly

[0086] Use modeling software to create a microfluidic chip model, make the above microfluidic chip with polymethyl methacrylate material, and seal it with acrylic glue. The openings of the microcolumn barriers in the microfluidic chip are 60, 40, and 20 μm respectively Take the photonic crystal microspheres of the above three sizes and modify them with antibodies to alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), and sugar chain antigen 199 (CA125), and pour the microspheres into the microfluidic chip before microfluidic control. The channel of the chip is pre-sealed with a blocking agent to prevent non-specific adsorption, and the microspheres are poured into the microfluidic chip, so that the microspheres are evenly mixed and arranged in the area corresponding to their size.

[0087] (2) Sample te...

Embodiment 3

[0090] The detection of serum markers of hepatitis B virus based on the microfluidic chip provided in Example 3

[0091] (1) Microfluidic chip assembly

[0092] The microfluidic chip model was created using SU8 glue lithography technology, and the above microfluidic chip was made of polydimethylsiloxane material, which was sealed with plexiglass by plasma, and the openings of the microcolumn barrier in the microfluidic chip were respectively 4.9, 3.0, and 1.0 μm, six microspheres of the above three sizes and two colors were respectively modified with hepatitis B virus surface antigen (HBSAg), hepatitis B virus surface antibody (HBSAb), hepatitis B virus Virus core antigen (HBcAg), hepatitis B virus core antibody (HBcAb), hepatitis B virus e antigen (HBeAg), hepatitis B virus e antibody (HBeAb). Before pouring the microspheres into the microfluidic chip, the channel of the microfluidic chip is pre-sealed with a sealing agent to prevent non-specific adsorption. area.

[0093]...

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Abstract

The invention discloses a multiple digital ELISA detection method and a microfluidic chip. The microfluidic chip includes a top layer chip, a microsphere area-arranged chip and microspheres with double-coded sizes and colors used in combination with the chip. The microsphere array chip is provided with microcolumn arrays arranged in parallel with different radius and size intervals. The microcolumn array of each size is the capture area of ​​microspheres of corresponding size, and the surface of the microspheres is the solid phase surface for detection. Size and color double-coded microspheres are constructed by mixing and superimposing different radius sizes and different ratios of two colors to construct high-throughput dual-classification coded microspheres. The surface of the microsphere is the solid phase surface for detection, which is double-coded by size and color, and the characteristics of the color are obtained by mixing different colors in different proportions. Each microsphere corresponds to a target, realizing large-scale simultaneous detection of multiple targets, and solving the problems of large sample and labor consumption, high time and economic costs in single target detection.

Description

technical field [0001] The invention relates to the field of rapid detection of medical devices, in particular to a multiple digital ELISA detection method and a microfluidic chip. Background technique [0002] On-site rapid testing, also known as instant testing, is an immediate testing and analysis performed at the patient's bedside or at the sampling site. Demand for on-site rapid detection for public health events or disease screening. [0003] The diagnosis of disease and the judgment of curative effect require joint detection and analysis of multiple targets (including genes or proteins, etc.), which poses new challenges to the rapidity and comprehensiveness of traditional testing fields. [0004] Single target detection consumes a lot of samples, takes a long time, and the cost of labor and reagents is high, which cannot meet the needs of large-scale screening and diagnosis of clinical diseases. However, the detection throughput of existing multiple detection methods...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/58G01N33/574G01N33/576G01N33/68B01L3/00
CPCG01N33/54313G01N33/582G01N33/57488G01N33/5762G01N33/5764G01N33/5761G01N33/6854B01L3/5027
Inventor 张元庆林谆董丽静
Owner 深圳市科瑞达生物技术有限公司