Nucleic acid detection method based on microfluidic microbead array chip
A technology of microbead array chip and detection method, which is applied in the field of biological detection, can solve the problems of low sensitivity, complicated detection process, difficult to realize micro sample analysis, etc., and achieve the effect of avoiding pollution
Active Publication Date: 2013-10-02
HUNAN INSTITUTE OF ENGINEERING
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[0004] The purpose of the present invention is to provide a new high-performance detection method that can be used for nucleic acid analysis, that is, a nucleic acid detection method based on a microfluidic microbead array chip, so as to solve the problems of low sensitivity and complicated detection process in current nucleic acid analysis methods. and difficult to achieve micro-sample analysis, and promote the further development of this technical field
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[0019] The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.
[0020] see figure 2 , is a schematic diagram of the principle of the method of the present invention. figure 2 The principle shown is as follows:
[0021] First, biotin-modified capture probe 3 and biotin-modified electron-rich protein 2 are modified to the surface of avidin-modified microspheres 1 according to a certain ratio to prepare functionalized microspheres 4 that can be used for nucleic acid detection; The target nucleic acid sequence 6 and the nanoparticles 5 modified with horseradish peroxidase (HRP) and detection probes are hybridized with the constructed functionalized microsphere array, and both the capture probe and the detection probe can hybridize with the target nucleic acid fragment, Form a sandwich structure 7; then flow into the enzyme-catalyzed substrate solution, including biotinylated tyramine 8 and hydrogen peroxid...
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The invention discloses a nucleic acid detection method based on a microfluidic microbead array chip. The method mainly comprises the steps that a functionalized microsphere is formed by fixing a specificity capture probe and electronic enriching protein on the surface of a microsphere, and is fixed in a microfluidic chip detection area; horse radish peroxidase and a detection probe are modified onto a nano particle to serve as signal labels; a target nucleic acid sequence and the nano particle flow into a detection area of the microfluidic microbead array detection chip to react with a microbead array; by a multienzyme nano signal amplification technology, biotinylated tyramine is bonded with the electronic enriching protein on the surface of the microsphere, and eluted, and then flows into a quantum point marked by avidin; the quantum point can identify and be fixed with biotin bonded with the electronic enriching protein on the surface of the microsphere; and quantitative detection is performed by fluorescence. The method can promote further development of the technical field, and solve the problems that the sensitivity is low, a detection flow is complicated and trace sample analysis is difficult to achieve in the current nucleic acid analysis method.
Description
technical field [0001] The invention belongs to the technical field of biological detection of a micro-total analysis system, and in particular relates to a nucleic acid detection method based on a microfluidic microbead array chip. Background technique [0002] Nucleic acid analysis is widely used in the field of biomedicine. At present, the main nucleic acid analysis methods include: microarray technology, polymerase chain reaction, Southern blot, chemiluminescence, fluorescence photometry, resonance light scattering, etc.; There are generally defects such as low sensitivity, expensive instruments, complicated detection process, and difficulty in multi-target parallel detection. Microfluidic bead array chip is a new chip model developed in the field of microfluidic chip research. It organically combines traditional microarray chips and microfluidic chips, and uses functionalized microspheres as sensing elements to realize High-throughput detection, using microfluidics as ...
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Inventor 张何傅昕刘涟朱振军
Owner HUNAN INSTITUTE OF ENGINEERING