Droplet-based microfluidic chip for detecting nucleic acid in high sensitive manner and using method thereof

Abstract

The invention discloses a droplet-based microfluidic chip for detecting nucleic acid in a high sensitive manner and a using method thereof. The droplet-based microfluidic chip has the front side and the back side, and comprises a sample feeding chamber, an oil phase sample feeding chamber and a second accommodating space, wherein samples pass through a microfluidic channel from the sample feedingchamber and is converged to the oil phase sample feeding chamber and the microfluidic channel; droplets are prepared and then flow into a first accommodating space; the droplets flow into the second accommodating space through a tilting chamber inlet from the first accommodating space. According to the droplet-based microfluidic chip for detecting nucleic acid in the high sensitive manner and theusing method thereof disclosed by the invention, functional areas are partitioned, three steps, including preparation of the droplets, PCR (Polymerase Chain Reaction) amplification and tilting statistical counting, preparation and amplification of the droplets, of the digital PCR are separately carried out in an individual area, micro-droplets are located in a free state, and the stability of themicro-droplets is high, so that the stability of the droplets is effectively ensured, an uncovering step of the digital PCR chip in a using process is avoided, and the crossed pollution risk is avoided, and thus, the success rate and the timeliness of implementation of the digital PCR are improved.

Description

technical field [0001] The invention relates to the technical field of sample analysis test tubes used in the detection or analysis of biological reactions, and in particular, to a droplet microfluidic chip for highly sensitive detection of nucleic acids and a method for using the same. Background technique [0002] Nucleic acid is one of the most basic substances of life, and any disease can find evidence at the nucleic acid level. At present, nucleic acid detection has been widely used in disease research, in vitro diagnosis, veterinary medicine, food safety, biosafety, agriculture and other fields. [0003] Currently, there are three methods for quantitative detection of nucleic acid molecules: photometric method, real-time fluorescence quantitative PCR and digital PCR (Digital PCR, dPCR) technology. Digital PCR technology is a nucleic acid quantification based on single-molecule PCR method for counting, and is a technology for nucleic acid molecule amplification and abs...

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Problems solved by technology

However, this limitation will increase the selectivity and difficulty of the process, resulting in increased processing costs

(d) During the digital PCR amplification process, it is difficult to maintain the state of micro-droplets tiled into a layer

The amplification process involves temperature cycling (the temperature varies from 4 to 96 degrees Celsius). The temperature change will induce the flow and the generation of air bubbles, disturb the state of the droplet tiling, and lead to the failure of digital PCR detection.

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