Droplet for multiple polymerase chain reaction (PCR) detection

Abstract

The invention discloses a droplet for multiple polymerase chain reaction (PCR) detection. The droplet comprises general content, a surface and a characteristic substance, wherein the general content is distributed inside the droplet and provides reaction conditions for the PCR; the exterior of the droplet is coated with the surface to form a stable droplet interface; the characteristic substance is positioned inside the droplet and is a solid material compatible with a biological reaction, a template molecular chain is immobilized on the surface of the solid material, and the template molecular chain can be used for realizing qualitative detection or quantitative determination of nucleic acid molecules by virtue of PCR amplified signals. According to the scheme, each droplet is similar toa tube of monoplex-PCR, the amplified reaction of each droplet is relatively independent, the problem that mutual interference exists because the traditional multiple PCR system is large can be effectively improved, and the droplet for multiple PCR detection is a very effective nucleic acid amplification method.

Description

technical field [0001] The invention belongs to the field of molecular detection, and in particular relates to a liquid droplet used for multiple polymerase chain reaction detection. Background technique [0002] Polymerase Chain Reaction (PCR) technology is currently the most widely used nucleic acid detection technology, and its main advantages are simplicity, stability, and good application foundation. The principle of multiplex PCR is to add multiple pairs of specific primers into the reaction system at the same time to amplify multiple DNA fragments at the same time. Since there are obvious differences in the size of the amplified fragments of each primer, according to the fragment size of the product or by molecular hybridization Realize simultaneous detection of multiple microorganisms. Since the concept of multiplex PCR was first proposed in 1988, after years of development, multiplex PCR technology has gradually matured. [0003] However, the main limitation of PC...

AI Technical Summary

Problems solved by technology

Existing multiplex detection techniques are firstly subjected to multiplex PCR amplification. This multitarget amplification has the following problems: unsatisfactory specificity and sensitivity; limited amplification of a specific fragment; For primers, the probability of forming primer-dimers increases; if non-specific products are amplified preferentially, a large amount of reaction substrates will be consumed and specific amplification efficiency will be reduced

At present, the detection methods of multiplex PCR mainly include capillary electrophoresis (or gel electrophoresis), liquid phase chip, mass spectrometry, etc. No matter which method is used for detection, the quality of multiplex amplification in the early stage will directly affect the detection results

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