A multifunctional integrated microfluidic nucleic acid analysis chip and its preparation and analysis method

Abstract

The invention provides a multifunctional integrated microfluidic nucleic acid analysis chip. The chip is prepared by preparing a combination mould according to 3D printing or micromachining technology, generating a microfluidic substrate according to an injection moulding method, and completing bonding of multiple layers including a chip substrate, a liquid circuit layer, an elastic film, a gas circuit layer and the like by means of plasma cleaning to finally obtain the complete chip. The chip integrates various structures including reagent cavities, mixing cavities, a splitting cavity, a DNA (deoxyribonucleic acid) extraction cavity, amplification reaction cavities, a microvalve, a microchannel and the like and has functions of sample loading, reagent mixing, heating for splitting, DNA adsorption, cleaning, eluting, amplification reaction and the like. The multifunctional integrated microfluidic nucleic acid analysis chip has the advantages that totally-closed automatic operation of 'sample input-result output' is realized, complicated manual operations are avoided, and detection efficiency is improved.

Description

technical field [0001] The invention relates to the technical field of microfluidic nucleic acid analysis chips, in particular to an integrated microfluidic nucleic acid analysis chip with multiple functions such as sample pretreatment, nucleic acid extraction and purification, and nucleic acid amplification, and a preparation and analysis method. Background technique [0002] As the basic genetic material, nucleic acid carries all kinds of information, and the understanding of its structure and function is beneficial to the study of genetics, evolution and disease diagnosis of species. Microfluidic chips have extensive and extremely important applications in nucleic acid analysis, such as gene mutation detection, pathogen gene identification, and food safety inspection. Although traditional PCR technology can realize DNA amplification and detection, it requires manual complex experimental operations, such as sample pretreatment, nucleic acid extraction, DNA amplification an...

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Problems solved by technology

The integrated nucleic acid extraction chip reported in this paper is aimed at blood lysate. Nucleic acid extraction and PCR reactions are performed on the chip, and blood samples cannot be tested directly.

"A microfluidic chip integrating DNA extraction and real-time PCR for the detection of bacteria in saliva" published by Emily A.Oblath et al. on Lab on a chip. Although the microfluidic chip in this article can realize direct processing of samples, However, the aluminum oxide film is fragile and the chip is difficult to manufacture, and the chip is an open-cavity design, which is easy to cause aerosol pollution and affect the accuracy of the test results.

"Integrated glass microdevice for nucleic acid purification, loop-mediated isothermal amplification, and online detection" published by Wu Qingqing et al. on Analytical chemistry. The chip in this article is an integrated chip that integrates nucleic acid extraction, loop-mediated isothermal amplification, and online analysis. , but the chip uses the micro-column method to extract nucleic acid, the extraction efficiency and the reliability of the micro-column need to be further studied, there is no integrated lysis function on the chip, the sample needs to be manually lysed and then added to the chip, and it can only be used for isothermal amplification. Conditions are limited

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