Droplet PCR amplification detection device based on microfluidic chip

Abstract

The invention discloses a droplet PCR amplification detection device based on a microfluidic chip. The device comprises the droplet microfluidic chip, an XYZ motion unit, a PCR amplification unit anda detection unit. A droplet containing a DNA molecule is introduced into the droplet microfluidic chip through a pipette after being generated, the chip is placed in the droplet PCR amplification unitto perform a PCR amplification reaction, and fluorochrome in the droplet is motivated through an LED lamp and light filter in the droplet detection unit, a camera is controlled to take a picture of the fluorescent droplet, the picture of the fluorescent droplet is focused through an autofocus algorithm and the control of a Z motion platform, and acquiring of images of different portions of the chip is completed through chip marking and the control of an XY motion platform; the captured images are spliced to obtain complete information of the chip droplet region images, thereby completing droplet PCR amplification detection. The device has the advantages that the operation is simple, the reagent consumption is low, the droplet stability is good, the detection accuracy is high, and the PCRamplification diagnosis and analysis can be efficiently completed.

Description

technical field [0001] The invention relates to the field of life medical detection and diagnosis, in particular to a detection device integrating droplet PCR amplification and droplet detection based on a microfluidic chip. Background technique [0002] Before the traditional PCR amplification technology, the sample is microdropletized, that is, the reaction system sample is dispersed into a large number of droplets with a diameter of micron, so that each droplet contains one or no target gene, and then PCR amplification , if there is a fluorescent signal, it is interpreted as 1, and if there is no fluorescent signal, it is 0, and then the initial copy number and concentration of the target gene can be obtained according to the ratio of the Poisson distribution to the number of positive droplets. This new nucleic acid detection and quantification method is Micro-droplet digital PCR technology. Compared with traditional amplification techniques, micro-droplet PCR does not d...

AI Technical Summary

Problems solved by technology

[0003] The general droplet PCR method often involves many manual steps, for example, the droplet is generated in a certain way (such as the cross method) and then introduced into the PCR tube, and then the PCR tube is placed in the PCR instrument for reaction amplification. Then import the chip in a certain way, and finally put the chip under a microscope to observe the fluorescent droplets. This method is not conducive to the automation of gene molecular diagnosis, the process is cumbersome, and there will be loss of reagents in the process of importing and exporting chips and PCR tubes many times. , which reduces the stability of the droplet and affects the accuracy rate; at the same time, the general droplet PCR amplification technology cannot observe the fluorescence of the droplet during the PCR amplification process in real time, and the droplet PCR amplification detection device based on the microfluidic chip Combining droplet reaction amplification and real-time detection, there is no need for frequent import and export processes to consume reagents and reduce accuracy. It has a high degree of automation, good amplification efficiency, low error probability, simple operation, and real-time recording of PCR amplification. Process droplet fluorescence

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