Nucleic acid detection microsphere and preparation method thereof, kit and high-throughput nucleic acid detection method

Abstract

The invention provides a nucleic acid detection microsphere and a preparation method thereof, a kit and a high-throughput nucleic acid detection method. The nucleic acid detection microsphere is composed of nuclear bodies with fluorescence coding information and a coating layer of primers or probes. A variety of the nucleic acid detection microspheres are randomly distributed into micro-droplets generated by a nucleic acid reaction solution of a to-be-detected sample. When a temperature rises to 60 DEG C or above, the coating layers melt, and the primers and the probes are released into the nucleic acid reaction solution of the to-be-detected sample, so a complete nucleic acid amplification reaction system is formed; and the fluorescence coding nuclear bodies are left in a nucleic acid amplification solution and serve as fluorescence labels for marking the micro-droplets. By detecting the nuclear bodies with fluorescence coding information in each micro-droplet, the micro-droplet withonly one nuclear body is screened out as an effective droplet and subjected to subsequent analysis to obtain primer and probe types in the effective micro-droplet; and then a report fluorescence signal after a nucleic acid amplification reaction is read so as to determine whether corresponding to-be-detected target nucleic acid molecules exist in the effective micro-droplet or not.

Description

technical field [0001] The present application relates to the field of nucleic acid detection and analysis, in particular to a nucleic acid detection microsphere, a preparation method, a kit and a high-throughput nucleic acid detection method. Background technique [0002] In recent years, PCR (Digital PCR, dPCR) technology has developed rapidly. Polymerase Chain Reaction (Polymerase Chain Reaction, PCR) is a molecular biology technique used to amplify specific DNA fragments. PCR includes droplet PCR detection method and chip detection method. [0003] The microdroplet PCR system performs microdroplet treatment on the sample before the traditional PCR amplification, that is, the reaction system containing nucleic acid molecules is divided into thousands of picoliter or nanoliter microdroplets, in which each microdroplet may not Contain nucleic acid target molecules to be detected, or contain one to several nucleic acid target molecules to be detected. After PCR amplificat...

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Problems solved by technology

[0004] Based on this, it is necessary to provide a method for high-throughput nucleic acid detection and analysis that can achieve high throughput, high sensitivity, and short detection time in response to the problems of repeated detection, heavy workload, and time-consuming traditional PCR detection technology. Nucleic acid detection microspheres, preparation method, kit and high-throughput nucleic acid detection method

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