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Micro-fluidic chip detection method based on magnetic bead technology and reagent freeze-drying technology and micro-fluidic chip

A microfluidic chip and magnetic bead technology, which is applied in chemical instruments and methods, fluid controllers, biological testing, etc., can solve the problems of large coefficient of variation in repeatable testing, poor chip practicability, and complex chips, etc., to reduce transportation and Effects of storage cost, quick release and mixing

Pending Publication Date: 2020-04-10
四川微康朴澜医疗科技有限责任公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the disadvantages are that due to various reasons such as chromatography methods, the coefficient of variation (CV) of the repeatability test is large, and the purpose of accurate quantification cannot be achieved.
[0004] The existing microfluidic chips on the market have the following disadvantages: 1) The chip is too complicated and the cost is too high; 2) The detection equipment is complicated, the detection time is long, and the throughput is low; 3) The chip is not practical
However, since this method measures the magnetic beads one by one, the precision and sensitivity of the instrument are very high, resulting in extremely high cost and cannot be applied in the POCT field.

Method used

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  • Micro-fluidic chip detection method based on magnetic bead technology and reagent freeze-drying technology and micro-fluidic chip
  • Micro-fluidic chip detection method based on magnetic bead technology and reagent freeze-drying technology and micro-fluidic chip
  • Micro-fluidic chip detection method based on magnetic bead technology and reagent freeze-drying technology and micro-fluidic chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0094] Example 1: Determination of Procalcitonin (PCT) by Double Antibody Sandwich

[0095] 1. Antibody labeling

[0096] 1. Magnetic bead labeling

[0097] Take 1 mg of COOH-magnetic beads, the diameter of which is 1 μm, and add it into 1 ml of 2-(N-morpholino)ethanesulfonic acid (MES) buffer solution with a pH of 6.0, and mix well. Then add 10 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and 10 mg of N-hydroxysuccinimide (NHS) and 0.1 mg of PCT capture antibody , vortexed and mixed at room temperature for 4 h. After the reaction was completed, the supernatant was removed by magnetic separation, and 1 ml of 50 mM glycine blocking solution with a pH of 8.5 was added, and after blocking for 1 hour, a magnetic bead-labeled PCT capture antibody was obtained.

[0098] 2. Fluorescent microsphere labeling

[0099] Take 1 mg of COOH-Eu-polystyrene fluorescent microspheres with a diameter of 300 nm, add them into 1 ml of MES buffer solution with a pH of ...

Embodiment 2

[0107] Embodiment 2: The concentration of T3 in the serum sample is determined by competitive method

[0108] 1. Antibody labeling

[0109] 1. Magnetic bead labeling

[0110] Take 1 mg of COOH-magnetic beads, the diameter of which is 3 μm, and add it into 1 ml of MES buffer solution with a pH of 6.0, and mix well. Add 10 mg of EDC, 10 mg of NHS and 0.1 mg of T3 capture antibody, and mix and react at room temperature for 4 h after vortexing. After the reaction was completed, the supernatant was removed by magnetic separation, and 1 ml of 50 mM glycine blocking solution with a pH of 8.5 was added. After blocking for 1 hour, the magnetic bead-labeled T3 capture antibody was obtained.

[0111] 2. Fluorescent microsphere labeling

[0112] Take 1 mg of COOH-Eu-polystyrene fluorescent microspheres with a diameter of 300 nm, add them into 1 ml of MES buffer solution with a pH of 6.0, and mix well. Then 10 mg of EDC, 10 mg of NHS and 0.1 mg of T3-BSA labeled antigen were added, vor...

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Abstract

The invention provides a micro-fluidic chip detection method based on a magnetic bead technology and a reagent freeze-drying technology and a micro-fluidic chip for realizing the method. According tothe method, on the basis of a microfluidic technology, mixing of a sample and a reagent, a flow mixing capture reaction, magnetic separation, cleaning, detection and the like are integrated on a chip,and all reagent components required by the reaction are integrated on the chip, so that the operation of a client is simple and convenient, and the detection can be completed anytime and anywhere. Aclosed micro-channel for a sample to flow is formed in the micro-fluidic chip, and the micro-channel comprises a mixing bin, a reaction bin, a delay channel, a capture bin and a waste liquid bin whichare sequentially communicated.

Description

technical field [0001] The invention relates to the technical field of microfluidic instant detection, in particular to a microfluidic chip detection method and a microfluidic chip based on magnetic bead technology and reagent freeze-drying technology. Background technique [0002] POCT (Point-of-care Testing), also known as point-of-care testing, or on-site rapid testing, refers to a testing method that is carried out at the sampling site and uses portable analytical instruments and supporting reagents to quickly obtain test results. As far as the current market situation is concerned, the detection system based on immunochromatography technology is the most widely used. This detection system is low in cost and easy to operate, so it is very popular in primary hospitals. However, the disadvantages are: the coefficient of variation (CV) of the repeatability test is large due to various reasons such as the chromatography method, and the purpose of accurate quantification can...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/50G01N35/00G01N21/64B01L3/00
CPCG01N33/54326G01N33/54313G01N33/50G01N35/00G01N21/6408B01L3/502769B01L3/50273B01L3/502753G01N2035/00158B01L2200/0647B01L2200/10B01L2300/0861B01L2300/0883B01L2400/043
Inventor 唐勇廖政
Owner 四川微康朴澜医疗科技有限责任公司
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