Preparation and application of electrochemical micro-fluidic device for nucleic acid isothermal amplification

Abstract

The invention provides preparation and application of an electrochemical micro-fluidic device for nucleic acid isothermal amplification. A micro-fluidic electrochemical sensor device is directly established on a printed electrode. The device comprises two layers of PDMS chip. The micro-channels on the upper layer are used to fully mix various samples. The channel area in the lower layer is divided into a reaction area and a detection area. A temperature-control element is integrated to the device. The electrode surface is subjected to functional modification. Corresponding primers aiming at the hepatitis B virus nucleic acid are designed. Electrochemical reporter molecules are grafted onto the primers. The labeled primers and the nucleic acid to be detected are evenly mixed, then the mixture is injected into a micro-fluidic chip, and a proper temperature is controlled to carry out isothermal amplification reactions on the nucleic acid sample. The labeled primers are combined to the electrode surface through the base pairing effect during the amplification process. Then the virus can be detected through the detection on the reporter molecules. The provided device has the advantages of miniaturization, low cost, and portability, can perform rapid and high-sensitive detection, and is convenient for diagnosis in outdoors / home.

Description

technical field [0001] The invention relates to a microfluidic device, in particular to a method for preparing an electrochemical microfluidic device for nucleic acid isothermal amplification and its application in medical detection. Background technique [0002] Microfluidic devices integrate or basically integrate the basic operation units involved in the fields of chemistry and biology such as sample transfer, separation and purification into a chip of several square centimeters (or even smaller), and form a network of microchannels. A technology platform that uses controllable fluid throughout the entire system to replace the various functions of conventional chemical or biological laboratories. Due to the advantages of low consumption, high throughput and miniaturization, this technology is widely used in clinical, diagnostic, pharmaceutical, and toxicological fields. It has great application potential in research and other fields. [0003] Electrochemical detection te...

AI Technical Summary

Problems solved by technology

[0004] At present, traditional microfluidic electrochemical POCT (even diagnosis) devices use complex magnetron sputtering technology, plastic rejection lithography technology, ion beam etching technology, wet etching technology, etc. to complete micro-electrodes on glass substrates. Preparation, usually a single-layer PDMS chip rectangular pipeline design; in a three-electrode system, if the materials used for the working electrode, reference electrode, and counter electrode are different, the electrode preparation process will be more complicated

The entire processing and preparation process needs to be completed in a clean room, which is costly. In addition, because the preparation process involves many links, and each link has a lot of parameter settings, the requirements for technical operators are very high, so it is difficult to guarantee the repeatability of different batches of products

At the same time, it is difficult to achieve mass production in this way

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