Quantitative analysis method and system for digital nucleic acid amplification based on micro-droplets

Abstract

The invention provides a method and a system for digital quantitative analysis of nucleic acid amplification based on micro-droplet. The method comprises the following steps: preparing a to-be-detected nucleic acid amplification reaction liquid which includes a to-be-detected nucleic acid template, a reaction buffer water solution, deoxyribonucleoside triphosphate, a primer, polymerase and a product marking substance; loading the prepared to-be-detected nucleic acid amplification reaction liquid in a micro-pipeline of which the two ends are both provided with an opening, wherein the micro-pipeline is arranged above an open container, and the open container contains an oily liquid containing a surfactant; enabling the opening of one end of the micro-pipeline to do up-and-down reciprocating vibration on the surface of a liquid in the open container or do leftward-and-rightward reciprocating vibration below the liquid surface so as to generate a plurality of droplets which are flatly laid at the bottom of the open container; performing nucleic acid amplification reaction on the plurality of droplets in the open container; acquiring product signals generated after the nucleic acid amplification reaction is ended, and performing quantitative analysis on the nucleic acid template. The digital nucleic acid amplification analysis method can be used for implementing quantitative detection on low-concentration nucleic acid substances in a micro-system and is simple and convenient to operate, high in efficiency and low in cost.

Description

technical field [0001] The present application relates to the technical field of nucleic acid quantitative analysis, in particular to a micro-droplet-based digital nucleic acid amplification quantitative analysis method and system. Background technique [0002] The traditional quantitative PCR (Polymerase Chain Reaction, Polymerase Chain Reaction) technology is based on the exponential amplification of nucleic acid templates during the reaction process, and the quantification of nucleic acid templates can be achieved by comparing the number of amplification cycles and analyzing the amount of PCR products after amplification . However, there are many factors that cause the traditional quantitative PCR method to have certain limitations and inaccuracies. For example, the amplification is not exponential at the beginning, which will affect the accuracy of the analysis method; Difficult to detect or can only be detected if the amplification is present twice or more, etc. [00...

AI Technical Summary

Problems solved by technology

However, the above-mentioned methods rely on relatively complex fluid drive devices, etc., which are costly and are not conducive to the application of digital nucleic acid amplification.

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