Microfluidic-based nucleic acid hybridization reaction platform and hybridization analysis method

Abstract

The invention provides a microfluidic-based nucleic acid hybridization reaction platform and a microfluidic-based nucleic acid hybridization analysis method. The platform consists of an upper layer and a lower layer, wherein the upper layer is a microfluidic chip controlled by a micropump and a microvalve; the lower layer is a substrate for fixing an oligonucleotide probe; and a fluid channel or a chamber is formed by the upper layer and the lower layer through irreversible sealing. The hybridization analysis method comprises a step of promoting liquid flow to automatically flow to and fro in a hybridization channel by taking positive pressure and negative pressure which are controlled by a program as a driving force, namely filling a sample into a microchannel by a capillary force or negative pressure after the sample enters a waste liquid chamber, and promoting liquid to flow to and fro in the microchannel by controlling the pushing and bounce (positive pressure and negative pressure) of two valve seats at both ends of the liquid channel. The microfluidic-based nucleic acid hybridization reaction platform and the microfluidic-based nucleic acid hybridization analysis method have the advantages that: mass transfer of materials is quick, hybridization time is short, the sample consumption is low, a hybridization signal of the sample can be detected in real time, the integration is easy to realize, a plurality of samples can be subjected to parallel analysis synchronously and the like.

Description

technical field [0001] The invention relates to nucleic acid hybridization analysis, in particular to a microfluidic-based nucleic acid hybridization reaction platform and a hybridization analysis method. Background technique [0002] DNA microarray (microarray) technology has become a very effective tool for drug screening, gene expression analysis and clinical diagnosis. The basic principle of microarray hybridization technology is that the DNA target molecules in the sample solution and the DNA probe molecules immobilized on the solid phase carrier undergo pairing hybridization according to the principle of base complementarity. However, this technology mainly relies on the diffusion of DNA target molecules in the solution, and due to the small diffusion coefficient and long diffusion distance of DNA molecules, the time to complete hybridization is very long (6-12h). [0003] In order to shorten the hybridization time, a variety of active or passive liquid mixing methods...

AI Technical Summary

Problems solved by technology

Continuous DNA hybridization mainly uses external drive equipment such as peristaltic pumps or centrifugal force to continuously flow the target molecule solution over the surface of the probe carrier to achieve rapid hybridization. A prominent disadvantage of this method is that the actual consumption is still relatively large compared with the reciprocating method; The reciprocating type is to make a certain volume (usually uL or nL) of the target molecule solution flow through the probe molecules repeatedly to achieve rapid hybridization.

Although a variety of methods have achieved low-cost and fast DNA hybridization, none of the methods can achieve the following functions and characteristics at the same time: 1. The automatic control of the hybridization process can be realized by using a micro-pump and micro-valve structure

Images