Microfluidic nucleic acid detection method and application

Abstract

The invention provides a microfluidic nucleic acid detection method. The method comprises the following steps: carrying out nucleic acid detection by using a disc-type microfluidic chip, wherein a reaction unit of the disc-type microfluidic chip comprises one or more sample adding pools, a micro-channel, a pressure balance channel and a reaction pool, and samples in the sample adding pools pass through the micro-channel to reach the reaction pool for reaction under the action of centrifugal force in the rotating process of the chip. The full-process nucleic acid detection including the steps of nucleic acid extraction, amplification, detection and the like can be automatically completed by an instrument after sample adding is completed. The method can be compatible with various different nucleic acid amplification methods such as loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA) and the like. 21 samples can be detected at the same time within 90 minutes, the detection sensitivity reaches 0.5 copy / microliter, and the method can be used for rapid pathogen infection screening in areas without large nucleic acid detection equipment such as villages and communities.

Description

technical field [0001] The invention belongs to the field of medical detection, and in particular relates to a microfluidic nucleic acid detection method and application. Background technique [0002] Microfluidic technology refers to the technology of manipulating tiny amounts of liquids in micron-scale pipes. Due to the unique fluid properties of fluids at the microscale, such as laminar flow and droplets, microfluidic technology can achieve operations that are difficult to achieve at the macroscale. Microfluidic chip, also known as lab-on-a-chip or micro-full analysis system, refers to the integration of sampling, dilution, reaction, separation, detection and other operation units required for analysis and detection on the same microchip. Compared with the traditional detection mode, the microfluidic chip has many advantages such as controllable liquid flow, less sample consumption, fast analysis speed, etc. New hot research fields. [0003] Nucleic acid is the genetic...

AI Technical Summary

Problems solved by technology

However, these detection devices still face the problems of complex equipment and high cost, and many devices only perform nucleic acid amplification and detection processes, and do not integrate nucleic acid extraction steps. The samples need to be cracked in advance and then added to the device for amplification. integrated automated nucleic acid detection

Although the existing technology can be used for nucleic acid extraction, amplification and detection, it still needs to manually manipulate the reaction liquid, which is automated and has a low degree of integration

And the existing technology has low throughput

The existing technology only integrates nucleic acid amplification and detection, but does not integrate nucleic acid extraction functions. Advanced nucleic acid extraction is required for detection, and the steps are cumbersome to use

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