Portable microfluidic PCR (Polymerase Chain Reaction) instrument and gene sample fluorogenic quantitative detection method

Abstract

The invention provides a portable microfluidic PCR (Polymerase Chain Reaction) instrument and a gene sample fluorogenic quantitative detection method. The portable microfluidic PCR instrument comprises a control unit, a PCR chip, a temperature control unit and a fluorescence signal acquisition unit, the temperature control unit and the fluorescence signal acquisition unit are connected with the control unit; the PCR chip comprises a substrate, a reaction chamber is formed on the end face of one side of the matrix, a sealing cover plate is covered on the reaction chamber, the reaction chamber is communicated with the outside through a capillary shaped sample inlet and a capillary shaped sample outlet on the substrate; the temperature control unit comprises a temperature detector, a heating part and a refrigerating part, the heating part and the refrigerating part heats or refrigerates the substrate in the PCR chip; an acquisition end of the fluorescence signal acquisition unit is distributed correspondingly to one side of the sealing cover plate of the PCR chip, so that a fluorescence image in the reaction chamber is acquired. The fluorescence signal acquisition unit instrument can conduct the polymerase chain reaction, meanwhile, the fluorescence signal acquisition unit acquires the reaction circulation fluorescence image, so that the fluorogenic quantitative detection of a gene sample is realized.

Description

Technical field [0001] The present invention relates to the field of biotechnology, in particular to a portable microfluidic PCR instrument. The invention also relates to a method for fluorescence quantitative detection of genetic samples based on the microfluidic PCR instrument. Background technique [0002] The basic principle of PCR (polymerase chain reaction) technology is similar to the natural replication process of DNA, and its specificity relies on oligonucleotide primers complementary to both ends of the target sequence. PCR basically consists of three basic reactions: denaturation-annealing-extension Step composition. PCR uses DNA denatured at a high temperature of 95°C in vitro to become single-stranded. At low temperatures (usually around 60°C), the primers and single-strands are combined according to the principle of base complementary pairing, and then the temperature is adjusted to the DNA polymerase for the most suitable reaction. At temperature (about 72°C), DNA...

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Problems solved by technology

In such a heating method, the amount of sample used is at least hundreds of microliters, which has a certain thermal inertia, and the equipment itself also has thermal inertia, so the frequency of temperature switching should not be too fast

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