Micro-fluidic chip immunodiagnosis kit and preparation method thereof

A microfluidic chip, immunodiagnosis technology, applied in measurement devices, instruments, scientific instruments, etc., can solve problems such as low capture efficiency, influence detection results, poor mixing effect, etc., to achieve shielding non-specific binding and detection accuracy. High, less liquid residue effect

Inactive Publication Date: 2019-04-19
TIANJIN RINGPU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although microfluidic chip technology currently has many advantages such as saving samples and detecting multiple antigens or antibodies at one time, the technology is still immature.
First of all, the miniaturization and portability of the microfluidic chip limit the size of the microchannel on the chip, which leads to the fact that the fluid in the chip can only be mixed by molecular diffusion, and the mixing effect is not good, which affects the sensitivity and adequacy of the reaction, and leads to detection The accuracy of the results is reduced; second, the existing microfluidic chips for immunodiagnosis often have poor antigen or antibody immobilization effects, which are mainly manifested in different batches of chips and the fixation of each point on the same chip. Uniformity difference, as well as non-specific adsorption lead to a decrease in the accuracy of the test results, and the low capture efficiency of the antigen or antibody on the target protein after immobilization affects the test results; in addition, the existing microfluidic chip antigen coating for immunodiagnosis It takes a long time, which indirectly affects the activity of the antigen or antibody after immobilization and the efficiency of capturing the target protein, thus affecting the sensitivity and accuracy of the detection results

Method used

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  • Micro-fluidic chip immunodiagnosis kit and preparation method thereof
  • Micro-fluidic chip immunodiagnosis kit and preparation method thereof
  • Micro-fluidic chip immunodiagnosis kit and preparation method thereof

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preparation example Construction

[0044] According to the second aspect of the present invention, a method for preparing a microfluidic chip immunodiagnostic kit includes the following steps: the microfluidic chip with fan-shaped depressions connected in series through tubular cavities treated sequentially with dopamine and glutaraldehyde The chip body is coated with antigen to obtain a microfluidic chip.

[0045] In a preferred embodiment, it includes the following steps: treating the dopamine-treated microfluidic chip body with fan-shaped depressions connected in series through tubular cavities with glutaraldehyde to obtain the microfluidic chip substrate, and then coating the antigen , to obtain a microfluidic chip.

[0046] In a preferred embodiment, the glutaraldehyde treatment includes the following steps: putting the dopamine-treated microfluidic chip body with a fan-shaped depression in series through a tubular cavity into glutaraldehyde with a volume fraction of 8-20% The absolute ethanol solution is...

Embodiment 1

[0055] Preparation of microfluidic immunodiagnostic kit:

[0056] A total of 10 untreated microfluidic chips with fan-shaped depressions connected in series through tubular cavities were prepared by etching, and each was designed with 6 sample injection holes.

[0057] Dopamine treatment: Soak the microfluidic chip body with 12mM Tris-HCl dopamine solution for 8 hours. During the immersion process, air was introduced into the system, washed with water for 3 times after immersion, and then air-dried naturally.

[0058] Glutaraldehyde treatment: Soak the dopamine-treated microfluidic chip body with 10% glutaraldehyde absolute ethanol solution at a room temperature of 25°C and keep it warm for 60 minutes, then take it out and wash it with water for 3 times. Then air-dried naturally to obtain the microfluidic chip substrate.

[0059]Coating antigen: add bovine foot-and-mouth disease type A virus, bovine foot-and-mouth disease type O virus, bovine foot-and-mouth disease Asian type...

Embodiment 2

[0061] Primary antibody: Blood was collected from experimental cattle by pricking method, and 500 μl of negative serum samples were prepared, and then immunized with homemade bovine epidemic diarrhea + bovine foot-and-mouth disease type A virus inactivated vaccine. 28 days after immunization, blood was collected by pricking method, and prepared Measure 500 μl of serum sample and dilute it 2500 times with PBS as the primary antibody.

[0062] Take the prepared microfluidic chip, and add 200 μl of serum to be tested, 800 μl of PBST cleaning solution, and 200 μl of 2000-fold diluted HRP-goat anti-bovine IgG into different sample wells.

[0063] Pre-set the automatic detector according to the following procedures, and perform the pumping according to the following procedures in sequence: pump the serum to be tested into the microfluidic chip and incubate at 25°C for 2 hours, then pump the PBST cleaning solution, wash 3 times, and pump out Cleaning fluid. Secondary antibody: pump ...

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Abstract

The invention provides a micro-fluidic chip immunodiagnosis kit and a preparation method thereof, and relates to the technical field of micro-fluidic chips. The micro-fluidic chip immunodiagnosis kitcomprises a micro-fluidic chip substrate and an antigen, the micro-fluidic chip substrate is provided with a sector-shaped concave portions formed by connecting tubular cavities in series, and the sector-shaped concave portions are treated with dopamine and glutaraldehyde; the number of the sector-shaped concave portions is larger than or equal to 4. The antigen covers the surfaces of part of thesector-shaped concave portions of the micro-fluidic chip substrate. According to the micro-fluidic chip immunodiagnosis kit, the technical problem that in the prior art, a micro-fluidic chip immunodiagnosis kit high in sensitivity and accuracy is in lack in the prior art is relieved; the preparation method of the micro-fluidic chip immunodiagnosis kit is provided. According to the method, antigencovering is fast, the process is simple, and the consumed time is short.

Description

technical field [0001] The invention relates to the technical field of microfluidic chips, in particular to a microfluidic chip immunodiagnostic kit and a preparation method thereof. Background technique [0002] A microfluidic chip is a technical product characterized by manipulating fluids in a micron-scale space, mainly through micro-electro-mechanical processing technology, on a chip of several square centimeters made of silicon, quartz, glass or polymer materials. A micro-pipeline network structure is built on top of the chip, and functions such as sampling, dilution, reagent addition, reaction, separation, and detection are concentrated on one chip, and controllable fluid flows in the chip to realize a multifunctional technology for conventional chemical or biological laboratories. platform. [0003] Although microfluidic chip technology currently has many advantages such as saving samples and being able to detect multiple antigens or antibodies at one time, the techn...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56905
Inventor 李守军付旭彬鲍恩东刘海霞蔡天宁王艳晓吕传忠
Owner TIANJIN RINGPU BIO TECH
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