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Magnetic particle direct chemiluminiscence microfluidic chip for whole blood sample testing

A microfluidic chip, whole blood sample technology, applied in chemiluminescence/bioluminescence, chemical instruments and methods, laboratory containers, etc., can solve the problems of interference, low sensitivity, long detection time, etc., and achieve high Sensitive detection, solving the effects of expensive instruments and high sensitivity

Active Publication Date: 2016-01-20
SHENZHEN HUAMAIXINGWEI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The technical problem to be solved by the present invention is to provide a microfluidic magnetic sensor for the problems of low sensitivity, poor repeatability, obvious interference, and the existing chemiluminescent microfluidic chip supporting equipment and long detection time of the existing rapid diagnostic method. The integrated chip of particle chemiluminescence immunoassay (integrating all components except the test sample into the chip) and supporting small portable equipment, so as to realize the rapid, accurate and highly sensitive quantitative detection of analytes in the field samples

Method used

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  • Magnetic particle direct chemiluminiscence microfluidic chip for whole blood sample testing
  • Magnetic particle direct chemiluminiscence microfluidic chip for whole blood sample testing
  • Magnetic particle direct chemiluminiscence microfluidic chip for whole blood sample testing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1: Determination of Procalcitonin (PCT) by Double Antibody Sandwich

[0073] (1) Antibody labeling

[0074] Add appropriate amount of acridinium ester, 10 μg EDC and 15 μg NHS solution and 10-30 μg anti-PCT monoclonal antibody solution to the phosphate buffer, mix well and react at room temperature for 4 hours, add 1 mg glycine to block. Separating and purifying with a chromatographic column or a chromatographic column to obtain an acridinium ester-labeled PCT antibody.

[0075] Add 1 mg magnetic particles (2 μm in size), 10 μg EDC and 15 μg NHS solution and 10-30 μg anti-PCT monoclonal antibody (different from acridinium ester-labeled antibody) solution to the phosphate buffer, mix well and react at room temperature for 2 hours, add 2 mg Glycine blocked. Magnetic adsorption purification to obtain magnetic particle-labeled PCT antibody.

[0076] (2) Microfluidic chip assembly

[0077] Acridine ester-labeled PCT antibody solution contains 0.1% bovine serum al...

Embodiment 2

[0086] Embodiment 2: Determination of tacrolimus blood drug concentration by competitive method

[0087] (1) Antibody / antigen labeling

[0088] Add an appropriate amount of magnetic particles (0.5 μm in size), glutaraldehyde and tacrolimus solution to the phosphate buffer, mix well and react at room temperature for 4 hours, then add glycine to block. Separation and purification with chromatographic column or chromatographic column to obtain magnetic particle-labeled antibody.

[0089] Avidin-labeled acridinium ester and biotinylated tacrolimus are mixed at a ratio of 1:0.5-1:2 to obtain acridinium-labeled tacrolimus.

[0090] (2) Microfluidic chip assembly

[0091] Acridine ester-labeled antigen solution contains 0.1% bovine serum albumin, 5% glycerol, 1% Triton X-100 and 0.01% Proclin300 pH 7.4 phosphate buffer solution; magnetic particle-labeled antibody solution contains 0.5% bovine serum Albumin, 0.1% casein, 0.2% Tween 20 and 0.01% Proclin300 in pH 7.4 phosphate buffer...

Embodiment 3

[0100] Example 3: Magnetic Particle Size Screening

[0101] For other experimental conditions, refer to Example 1. The size of the magnetic particles and the magnetic induction of the magnet are carried out according to the following scheme.

[0102] The particle size is 0.1 μm, 0.5 μm, 1.0 μm, 2.0 μm, 2.8 μm, 5 μm, 10 μm. The magnetic induction of the magnet is 500 Gauss, 1000 Gauss, 4000 Gauss, 8000 Gauss, 12000 Gauss, 30000 Gauss. Magnetic particles of seven sizes are respectively driven by the six kinds of magnets.

[0103] The experimental results show that when 0.1μm magnetic particles are combined with a 500 Gauss magnet, the minimum detection limit is 100pg / ml, the quantitative detection range is 0.1-100ng / ml, and the linear correlation coefficient R 2 >0.95; intra-assay and inter-assay repeatability are less than 20%. That is: the chemiluminescent signal is weak, the sensitivity is not high, and the repeatability is poor.

[0104] When 10μm magnetic particles and ...

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Abstract

The invention relates to a magnetic particle direct chemiluminiscence microfluidic chip for whole blood sample testing. The chip is composed of a top adhesive tape (10), a chip substrate (1) and a bottom adhesive tape (13); the chip substrate comprises a filtering area (2), a coating area (3), a cleaning area (4), a testing area (5), a cleaning liquid storing pool (7), a luminescence substrate liquid storing pool (8) and a waste liquid pool (11), wherein the testing area is connected with the cleaning liquid storing pool and the luminescence substrate liquid storing pool through liquid releasing channels (9) respectively; the top adhesive tape comprises a sample feeding opening (11) and a cleaning liquid storing pool demising hole (12).

Description

technical field [0001] The present invention relates to a chip that uses magnetic particle direct chemiluminescence technology and microfluidic chip technology to realize highly sensitive quantitative detection of analytes, and particularly discloses a magnetic particle direct chemiluminescence microfluidic chip for whole blood sample detection, It can be applied to biomedical research, clinical diagnosis, biochemical detection, and judicial identification, and belongs to the field of microfluidic chip chemiluminescence technology. Background technique [0002] At present, there are two main development trends for in vitro diagnostics (IVD): one is automation and integration, that is, the use of fully automated and highly sensitive large-scale instruments and equipment in the central laboratory of a large hospital to achieve high-precision disease analysis and diagnosis ; Another kind of miniaturization and bedside, that is, to realize rapid analysis and diagnosis on the spo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76B01L3/00
Inventor 王东李泉
Owner SHENZHEN HUAMAIXINGWEI MEDICAL TECH CO LTD
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