Multifunctional integrated microfluidic nucleic acid analysis chip and preparation and analysis method thereof

A nucleic acid analysis and microfluidic technology, applied in biochemical equipment and methods, biochemical instruments, biochemical cleaning devices, etc., can solve problems such as limited use conditions, inability to test blood samples, affecting the accuracy of test results, etc. simple effect

Active Publication Date: 2016-02-24
合肥中科易康达生物医学有限公司
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  • Summary
  • Abstract
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  • Application Information

AI Technical Summary

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.
"Amicrofluidic chip integrating DNA extraction and real-time PCR for the detection of bacteria insaliva" published by EmilyA.Oblath et al. on Labonachip. Although the microfluidic chip in this paper can realize direct processing of samples, the aluminum oxide film is fragile, making it difficult to make the chip, and the chip is an open-cavity design. It is easy to cause aerosol pollution and affect the accuracy of the test results
"Integrated glass microdevice

Method used

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  • Multifunctional integrated microfluidic nucleic acid analysis chip and preparation and analysis method thereof
  • Multifunctional integrated microfluidic nucleic acid analysis chip and preparation and analysis method thereof
  • Multifunctional integrated microfluidic nucleic acid analysis chip and preparation and analysis method thereof

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example 1

[0054] The combined mold of the liquid layer substrate and the mold of the air layer chip are injected with PDMS glue, heated and cured, and the substrate is removed. A microfluidic chip is bonded by plasma cleaning substrates, liquid path layer substrates, elastic films, gas path layer substrates, etc. At this time, the microfluidic chip is processed for biocompatibility, high temperature sterilization and other related processing required for the chip before the PCR reaction. The GAPDH internal reference gene is used for detection and analysis, and 20 uL of human whole blood is added to the reagent chamber 81, and placed on the supporting detection and analysis platform. First, the blood and the lysing reagent are mixed through the microchannel and the mixing chamber, and then enter the lysing chamber for lysing. The lysed lysate is mixed with ethanol into the extraction chamber, the DNA is adsorbed on the silica gel membrane, and the waste liquid is pumped into the waste l...

example 2

[0056] The chip can also detect pathogenic microorganisms in food, taking the detection of Salmonella in dairy products as an example. After the dairy products are pre-cultured by pre-enrichment, take a little bacterial solution and mix it with the lysate, put it into the cracking chamber of the chip for lysing, and then mix it with ethanol into the extraction chamber embedded with a silica gel membrane, and the waste liquid is pumped into the waste liquid chamber, followed by Washing solution is added to clean the nucleic acid adsorbed on the silica gel membrane, and after drying, the eluent is added to elute the DNA. Specific detection of conserved invasiveness-associated genes in Salmonella spp. using loop-mediated isothermal amplification (LAMP).

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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...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12M1/34
CPCC12Q1/6844C12Q1/686C12Q2545/113C12Q2531/119
Inventor 刘勇朱灵邓国庆王贻坤赵树弥朱灿灿
Owner 合肥中科易康达生物医学有限公司
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