Pathogen detection method based on micro-fluidic chip

A microfluidic chip, pathogen detection technology, applied in the fields of chemistry and biomedicine, can solve the problems of high cost, complex processing, time-consuming, etc., and achieve the effects of high specificity, strong maneuverability, and excellent optical properties

Inactive Publication Date: 2013-01-02
WUHAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These technologies have played a huge role in clinical diagnosis, but there are s

Method used

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  • Pathogen detection method based on micro-fluidic chip
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  • Pathogen detection method based on micro-fluidic chip

Examples

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Embodiment 1

[0026] (1) Manufacturing method of integrated chip

[0027] Using standard soft lithography production technology, a layer of AZ50XT photoresist is cast on the clean and flat surface of the silicon wafer to obtain a thickness of about 40 μm. Under the cover of the printed film mask, it is exposed and developed with ultraviolet rays to obtain A standard mask that protrudes from the surface of the silicon wafer. Fix two self-made magnetic rods on both sides of the male mold microchannel, the angle between the magnetic rods is 0-180 o , the angle between the plane formed by the magnetic bar and the plane of the microchip is 90 o , reconcile the prepolymer mass ratio of polydimethylsiloxane (PDMS) to 10:1 (mass ratio), vacuum out the air bubbles, pour it on the mask, and then bake it in an oven at 75°C for 3-4 hours, After curing, cut it with a scalpel, lift it from the mask, and copy the pattern to the PDMS, use a flat punched needle to punch the sample and sample outlet; clean...

Embodiment 2

[0036] An integrated chip was fabricated by using the method in Example 1, and magnetic balls modified with anti-H9N2 avian influenza virus surface protein HA monoclonal antibody and biotinylated anti-H9N2 avian influenza virus surface protein HA monoclonal antibody were prepared. Then a magnetic sandwich immunofluorescence reaction was performed on the chip to detect H9N2. Figure 4 Fluorescent images (a) and corresponding spectral images (b) of different concentrations of H9N2 magnetic fluorescence immunoassay. Figure 5 is the change trend diagram (a) of different concentrations of H9N2 and fluorescence intensity, and the standard curve diagram (b), and Figure 4 correspond. Figure 6 In order to detect the specificity analysis of the H9N2 magnetic sandwich immunofluorescence detection method, the fluorescence picture (a) and the corresponding spectrum picture (b). Figure 7 In order to detect the specificity analysis of H9N2 magnetic sandwich immunofluorescence detecti...

Embodiment 3

[0038] An integrated chip was fabricated by using the method in Example 1, and magnetic balls modified with anti-Newcastle disease virus (NDV) surface protein HA monoclonal antibody and biotinylated anti-Newcastle disease virus (NDV) surface protein HA monoclonal antibody were prepared. Then a magnetic sandwich immunofluorescence reaction was performed on the chip to detect H9N2. Figure 8 Fluorescence graphs, spectrograms, and standard curves of magnetic fluorescent immunoassays for different concentrations of NDV. Figure 9 In order to detect the specificity analysis of the NDV magnetic sandwich immunofluorescence detection method, it includes the fluorescence diagram, spectrum diagram and histogram of the reagent blank and negative control.

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Abstract

The invention relates to a pathogen detection method based on a micro-fluidic chip. The method adopts a micro-fluidic chip integrated with a micro-magnetic field as a reaction container, adopts magnetic balls as solid phase carriers, and adopts quantum dots modified by streptavidin (SA-QDs) as fluorescent markers; under the action of the micro-magnetic field, magnetic balls are captured at specific parts in a chip channel, and thus a micro-reaction zone is formed; pathogens are captured through a sandwich immunization reaction; with the interaction between biotin and streptavidin, the immunofluorescence quantitative analysis of the pathogens is realized. The detection method combines the micro-fluidic chip, magnetic immunoassay, and fluorescence detection, has the advantages of rapidness, high efficiency, and few sample using amount for the micro-fluidic chip, high specificity and strong manoeuvrability for magnetic immunoassay, and excellent optical properties for quantum dots, and is a multi-objective real-time pathogen detection method.

Description

[0001] technical field [0002] The invention relates to a pathogen detection method based on a microfluidic chip, belonging to the fields of chemistry and biomedicine. Background technique [0003] In recent years, there have been successive outbreaks of SARS, bird flu, hand, foot and mouth disease, and H1N1 flu. Every epidemic of serious infectious diseases has caused huge losses to society and economy, and has brought great threats to people's lives. Currently commonly used pathogen detection methods include PCR technology, culture technology, immunoenzyme technology (EIA), enzyme-linked immunosorbent assay (ELISA) and so on. These techniques have played a huge role in clinical diagnosis, but there are still some shortcomings, most of which are time-consuming, complicated and costly. Establishing a rapid, sensitive and specific method for detecting pathogens is an important guarantee for the prevention and control of infectious diseases. [0004] Microfluidic chip tech...

Claims

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Application Information

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IPC IPC(8): G01N33/53
Inventor 张志凌张瑞巧李安珺庞代文
Owner WUHAN UNIV
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