Micro-fluidic chip immunofluorescence rapid detection kit as well as preparation and detection methods thereof

A microfluidic chip and detection kit technology, applied in the field of immune detection, can solve the problems of inability to integrate the reaction, high detection cost, long time-consuming, etc., so as to save repeated sampling operations, less reagent consumption, and detection time. short effect

Pending Publication Date: 2017-08-15
沈阳微流控生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This patent has the following deficiencies: (1) The antigen and antibody reagents used still need to be stored in reagent bottles separately (same as the traditional method), and the liquid flow, mixing and reaction are driven by vacuuming during the test, and the reaction cannot be integrated; (2) The solid phase carrier used - polyester membrane instead of microporous plate (or test paper), the cost is high; (3) The pore size of polyester membrane affects the enrichment of antigen or antibody; (3) The heterogeneous reaction is carried out on the solid phase carrier The surface reacts, the reaction is insufficient, and the sensitivity

Method used

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  • Micro-fluidic chip immunofluorescence rapid detection kit as well as preparation and detection methods thereof
  • Micro-fluidic chip immunofluorescence rapid detection kit as well as preparation and detection methods thereof
  • Micro-fluidic chip immunofluorescence rapid detection kit as well as preparation and detection methods thereof

Examples

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Embodiment 1

[0054] Example 1: Preparation and detection of N-terminal pro-brain natriuretic peptide (NT-proBNP) microfluidic chip fluorescent immunoassay kit reagents (multiple samples, 1 index detection kit).

[0055] 1. Prepare plate A and plate B.

[0056] Prepare plate A and plate B, both of which contain 12 detection units corresponding to each other. For the connected channel, the detection unit on the B board contains the sample loading tank, the lower part of the fluorescent probe tank, the lower part of the reaction detection tank and the lower part of the waste liquid tank corresponding to the A board.

[0057] 2. Preparation of labeled antibody fluorescent microspheres.

[0058] Fluorescent microspheres with a diameter of 100nm polystyrene-wrapped quantum dots (Qd, fluorescent probe) were washed, sonicated, and stirred, and then coupled with NT-proBNP monoclonal antibody 1 to form a "labeled antibody" after the coupling reaction. 1-Fluorescent microspheres", adding BSA buffer...

Embodiment 2

[0069] Example 2: Preparation and detection of preoperative 4-item (hepatitis B surface antigen / hepatitis C antibody / human immunodeficiency virus antibody / Treponema pallidum antibody) joint inspection kit (3 samples, 4-item joint inspection kit).

[0070] 1. Preparation of labeled antibody (or antigen) fluorescent microspheres: take polystyrene-coated europium metal chelate (Eu, fluorescent probe) fluorescent microspheres with a diameter of 200 nm, wash, sonicate, and stir, and mix with ① Hepatitis B surface antibody 1 ② Hepatitis C recombinant antigen 1 ③ Human immunodeficiency virus recombinant antigen 1 ④ Treponema pallidum recombinant antigen 1 for coupling, and after the coupling reaction, 4 kinds of fluorescent microsphere markers were formed respectively for future use.

[0071] 2. Preparation of capture antibody (or antigen) microspheres: Take polystyrene magnetic microspheres with a diameter of 3um, wash, sonicate, and stir, and mix with ① Hepatitis B surface antibody ...

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Abstract

The invention discloses a micro-fluidic chip immunofluerescence rapid detection kit as well as preparation and detection methods thereof. The kit comprises a micro-fluidic chip; the micro-fluidic chip comprises one or more detection units; the detection units are arranged on a radius axis of the micro-fluidic chip and are arranged in a divergence manner by taking the micro-fluidic chip as a center point; and a sample feeding groove, a fluorescence probe groove, a reaction detection groove, a waste liquid groove and microchannels which are mutually connected are sequentially arranged on the detection units from the center point of the chip to the edge. By using the kit, the dosage of a reagent is small, the integration degree is high, and a reaction systems is a homogeneous phase; and all the reagents are solidified in the chip in advance, and antigen-antibody reaction is carried out in a relatively closed space, so that crossed contamination and environmental pollution do not occur, and the kit has the advantages of high efficiency, high flux, high sensitivity, rapidness, sensitiveness, stability and the like.

Description

technical field [0001] The invention belongs to the field of immunological detection, and in particular relates to a microfluidic chip fluorescence immunological rapid detection kit and a preparation and detection method thereof. Background technique [0002] In recent years, the technology of microfluidic chips has developed rapidly, and its applications in medicine, life science and other fields have been continuously expanded. Microfluidic chip refers to the use of microfabrication technology to integrate the microchannel network structure and other functional components on a substrate of several square centimeters, and to control the fluid in the microchannel to realize sampling, dilution, mixing, reaction, The micro-total analysis system with multiple functions such as separation and detection has the remarkable advantages of miniaturization, integration, fast analysis speed, and less reagent consumption. [0003] Immunolabeling technology is based on the basic princip...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N33/533G01N33/577G01N33/543G01N33/576G01N33/571G01N33/569
CPCG01N21/6428G01N33/533G01N33/54326G01N33/56983G01N33/571G01N33/5764G01N33/5767G01N33/577G01N2446/90
Inventor 李祝华白众瑜黄雪莹孙颖孙秒吕辉高占岩
Owner 沈阳微流控生物科技有限公司
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