Droplet digital PCR chip, detection system, detection method and manufacturing method

Abstract

The invention relates to a droplet digital PCR chip, a detection system, a detection method and a manufacturing method. The droplet digital PCR chip comprises a chip body, wherein the chip body is provided with a liquid injection pool, a droplet generation pool, a flow pool, a droplet storage pool and a first driving source, the liquid injection pool, the droplet generation pool, the flow pool andthe droplet storage pool are sequentially communicated, the first driving source is communicated with the liquid injection pool, and the bottom of the liquid injection pool comprises a diaphragm; thedroplet generation pool is provided with a droplet generation cavity; a sample injection opening is formed in one end of the liquid injection pool, the other end of the liquid injection pool extendsinto the droplet generation cavity, and a first capillary pipeline is formed in a communicating path of the liquid injection pool and the droplet generation cavity; and the first driving source can drive the diaphragm to elastically expand and contract so as to extrude liquid in the liquid injection pool out of the droplet generation cavity through the first capillary pipeline to form droplets. The chip can be directly subjected to PCR, secondary transfer of the droplets is not needed, the state of the droplets is better maintained, PCR is facilitated, the detection efficiency and accuracy areimproved, and the chip has a wide application prospect in droplet digital PCR devices.

Description

technical field [0001] The invention relates to the technical field of microfluidic droplet digital polymerase chain reaction, in particular to a droplet digital PCR chip, a detection system, a detection method and a manufacturing method. Background technique [0002] Droplet digital PCR is to use a microfluidic chip to perform droplet processing on samples before traditional PCR amplification, that is, the reaction system containing nucleic acid molecules is divided into thousands of nanoscale droplets, where each droplet or Does not contain nucleic acid target molecules to be detected, or contains nucleic acid target molecules to be detected. After PCR amplification, each droplet is detected one by one. The droplet with fluorescent signal is interpreted as 1, and the droplet without fluorescent signal is interpreted as 0. According to the Poisson distribution principle and the number and proportion of positive droplets, The starting copy number or concentration of the tar...

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Problems solved by technology

Among them, the micro-droplet size obtained in this way has the advantages of uniformity, stable interface, and high PCR reaction efficiency. However, since PDMS is used as the chip material, and the PDMS material has particularly good air permeability and water permeability, it can be used directly on this chip. Performing PCR will cause the PCR reaction solution to evaporate and cause inaccurate results

And the chip only has the function of droplet generation, but not the function of detecting the reaction product, so the function of the chip is not complete

In addition, the chip is also equipped with a single-phase droplet forming microstructure at the three reagent injection ports, and the structure is relatively complex.

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