Integrated digital nucleic acid amplification chip based on micro-droplets as well as use method and application of integrated digital nucleic acid amplification chip

Abstract

The invention provides an integrated digital nucleic acid amplification chip based on micro-droplets as well as a use method and an application of the integrated digital nucleic acid amplification chip. The integrated digital nucleic acid amplification chip comprises an upper cover and a bottom plate. The upper cover comprises an oil inlet, a sample inlet and an exhaust port. The bottom plate comprises a droplet generation area and a droplet storage area, wherein the droplet generation area comprises an oil inlet flow channel and a sample inlet flow channel; the oil inlet flow channel is connected with the sample inlet, the sample inlet flow channel is connected with the sample inlet, the intersection of the oil inlet flow channel and the sample inlet flow channel is a cross-shaped channel, and micro-droplets are generated at the intersection of the oil inlet flow channel and the sample inlet flow channel; the droplet storage area comprises a collection cavity flow channel and a droplet storage cavity; the collection cavity flow channel is connected with the cross-shaped channel, and the droplet storage cavity is connected with the collection cavity flow channel and the exhaust port. The processes of droplet generation, nucleic acid amplification, detection and the like can be completed on the chip, so that the detection efficiency and the detection accuracy are improved, and rapid development of an integrated digital nucleic acid amplification technology based on micro-droplets is promoted.

Description

technical field [0001] The invention belongs to the field of nucleic acid detection, and in particular relates to an integrated digital nucleic acid amplification chip based on micro-droplets and its use method and application. Background technique [0002] At present, in vitro diagnostic technology using nucleic acid as a detection marker has received more and more attention, and has been widely used in clinical diagnosis, food safety, public health and other fields. Especially for the recent global outbreak of novel coronavirus pneumonia (COVID-19) caused by a new type of coronavirus infection, nucleic acid testing plays an important role in differential diagnosis, clinical guidance, discharge screening, etc. powerful means. [0003] In addition, for the emerging real-time detection application scenarios that do not depend on the central laboratory, the development of portable, low-cost, and integrated nucleic acid detection devices is of great significance, and it will h...

AI Technical Summary

Problems solved by technology

[0007] Among the existing commercialized digital PCR chips, most of them use one chip to generate droplets, transfer them to the PCR tube for amplification, and then transfer them to another detection chip for fluorescence detection, that is, the entire reaction is performed by "generated droplets-PCR Heating reaction-detection consists of three parts, and the operation is relatively cumbersome. At the same time, in the process of droplet transfer, there are droplet breakage, incomplete transfer, and aerosol cross-contamination, which reduces the accuracy and reliability of digital PCR.

Currently, there are relatively few integrated digital nucleic acid amplification chips (including digital PCR and digital isothermal amplification) based on micro-droplets. Thus affecting the detection accuracy