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Magnetic particulate chemiluminiscence micro-fluidic chip capable of quantitatively detecting brain natriuretic peptide in whole blood

A microfluidic chip, quantitative detection technology, applied in chemiluminescence/bioluminescence, analysis through chemical reaction of materials, biological testing, etc., can solve the problems of low sensitivity, interference, expensive chemiluminescence supporting equipment, etc. Achieve the effect of high sensitivity and high sensitivity detection

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 method for quantitatively detecting brain natriuretic peptide in whole blood for the problems of low sensitivity, poor repeatability and obvious interference of the existing rapid diagnostic method, as well as the expensive and long detection time of the existing chemiluminescence supporting equipment. The magnetic particle chemiluminescent microfluidic chip (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 BNP in the whole blood sample on site

Method used

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  • Magnetic particulate chemiluminiscence micro-fluidic chip capable of quantitatively detecting brain natriuretic peptide in whole blood
  • Magnetic particulate chemiluminiscence micro-fluidic chip capable of quantitatively detecting brain natriuretic peptide in whole blood
  • Magnetic particulate chemiluminiscence micro-fluidic chip capable of quantitatively detecting brain natriuretic peptide in whole blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1: Enzymatic Chemiluminescent Determination of BNP

[0070] (1) Antibody labeling

[0071] Add appropriate amount of HRP, 10 μg EDC, 15 μg NHS solution and 10-30 μg anti-BNP monoclonal antibody solution to the phosphate buffer, mix well and react at room temperature for 4 hours, add 1 mg glycine to block. Separation and purification with chromatographic column or chromatographic column to obtain HRP-labeled BNP antibody.

[0072] Add 1 mg magnetic particles (2 μm in size), 10 μg EDC and 15 μg NHS solution and 10-30 μg anti-BNP monoclonal antibody (different from HRP-labeled antibody) solution to the phosphate buffer, mix well and react at room temperature for 4 hours, add 1 mg glycine to block . Magnet adsorption enrichment and purification to remove unreacted anti-BNP antibody to obtain magnetic particle-labeled BNP antibody.

[0073] (2) Microfluidic chip assembly

[0074] HRP-labeled BNP antibody solution contains 0.1% bovine serum albumin, 0.1% Tween 20 ...

Embodiment 2

[0083] Embodiment 2: direct chemiluminescence assay BNP

[0084] (1) Antibody labeling

[0085] Add an appropriate amount of activated acridinium ester and 10-30 μg anti-BNP monoclonal antibody solution to the phosphate buffer, mix well and react at room temperature for 4 hours, and add 1 mg glycine to block. Separating and purifying with a chromatographic column or a chromatographic column to obtain an acridinium ester-labeled BNP antibody.

[0086] Mix avidinized magnetic particles (with a particle size of 0.5 μm) and biotinylated BNP antibody at a ratio of 1:1000 to 1:100000 to obtain magnetic particle-labeled BNP antibody.

[0087] (2) Microfluidic chip assembly

[0088] The acridinium ester labeled antibody solution contains bovine serum albumin, Tween-20 and Proclin300 pH7.4 phosphate buffer solution; the magnetic particle labeled antibody solution contains bovine serum albumin, casein, sucrose, Tween-20, Qular Pass X-100 and Proclin300 pH7.4 phosphate buffer.

[008...

Embodiment 3

[0097] Example 3: Magnetic Particle Size Screening

[0098] Refer to Example 2 for other experimental conditions, and the magnetic particle size and magnetic induction of the magnet are carried out according to the following scheme.

[0099] The particle size is 0.1 μm, 0.5 μm, 0.7 μm, 1.6 μm, 2.4 μm, 3 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.

[0100] The experimental results show that when 0.1μm magnetic particles and 500 Gauss magnets are used in combination, the minimum detection limit is 30pg / ml, the quantitative detection range is 30-4000pg / ml, and the linear correlation coefficient R 2 >0.93; intra-assay and inter-assay repeatability are both less than 20%. That is: the chemiluminescent signal is weak, the sensitivity is not high, and the repeatability is poor.

[0101] When 10μm magnetic particles ...

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Abstract

The invention discloses a magnetic particulate chemiluminiscence micro-fluidic chip capable of quantitatively detecting brain natriuretic peptide (BNP) in whole blood. The micro-fluidic chip is composed of top adhesive tape (12), a chip substrate (1) and bottom adhesive tape (15), wherein a filtering zone (2), a magnetic particle tagged BNP antibody coating zone (3), a reaction zone (5), a cleaning zone (6), a detection zone (7) and a liquid release channel (8) which are arranged on the chip substrate (1) are sequentially connected; a tagged BNP antibody storage tank (4) on the chip substrate (11) is connected with the reaction zone (5), and the detection zone (7) is connected with a cleaning liquid storage tank (9) and a luminous substrate liquid storage tank (10) through the liquid release channel (8).

Description

technical field [0001] The invention relates to a method for realizing high-sensitivity quantitative detection of BNP by using magnetic particle chemiluminescence technology and microfluidic chip technology, and particularly discloses a magnetic particle chemiluminescence microfluidic chip for quantitative detection of whole blood midbrain natriuretic peptide, which can The invention provides accurate and highly sensitive quantitative detection of BNP in whole blood, and belongs to the technical field of microfluidic chip chemiluminescence immunoassay. Background technique [0002] Brain natriuretic peptide is a polypeptide with strong natriuretic, diuretic, vasodilator and antihypertensive effects isolated from pig brain for the first time in 1988 by Japanese scholar Tetsuji Sudoh. A large number of basic and clinical studies have shown that the level of BNP in the blood increases significantly in heart failure. As a new biomarker, it is of great value in the diagnosis of h...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/68
Inventor 王东李泉
Owner SHENZHEN HUAMAIXINGWEI MEDICAL TECH CO LTD
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