Real-time fluorescence PCR (polymerase chain reaction) detection system based on rotary type microfluidic chip

Abstract

The invention relates to a real-time fluorescence PCR (polymerase chain reaction) detection system based on a rotary type microfluidic chip. The system comprises an annealing low-temperature area, an extension appropriate temperature area, a thermal change high temperature area, a rotating module, a PCR reagent accommodating cavity, an injection micro-channel, an instrument fixing base and a fluorescence detection system, wherein a temperature cycle control module comprises the annealing low-temperature area, the extension appropriate temperature area and the thermal change high temperature area; a micro-channel cycle PCR amplification module comprises the rotating module, the PCR reagent accommodating cavity, the injection micro-channel and the instrument fixing base; and a fluorescence spectrum detection module comprises the fluorescence detection system. According to the system, three modules, namely, the micro-channel cycle PCR amplification module, the temperature cycle control module and the fluorescence spectrum detection module are integrated, so that the portable micro PCR fluorescence real-time detection system applicable to space operation requirements is realized, and purposes of function integration, structure microform, lightness, small size and full-automatic detection are achieved.

Description

technical field [0001] The invention relates to a real-time fluorescent PCR detection system based on a rotary microfluidic chip, belonging to the fields of biology, analytical chemistry and medical detection. Background technique [0002] At present, in space vehicles during long-term space flight, the growth of microbial flora and strain variation will affect the performance of instruments and equipment and working environment conditions, and cause harm to the normal operation of spacecraft and its instruments and equipment. In order to prevent the occurrence, spread and effective treatment of infectious diseases in space flight, a real-time automatic alarm system for space on-orbit biological hazards is urgently needed, and its core is a microbial nucleic acid fluorescence detection microsystem. Due to the space on-orbit, the equipment is required to have the technical characteristics of full automation, small volume and small weight. Therefore, simplifying the microflui...

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Problems solved by technology

[0005] In the current market, common real-time fluorescent PCR detection instruments cannot meet the needs of the space field, and mainly have the following shortcomings:

[0006] 1. The temperature control module in the PCR amplification system in the instrument uses a metal block for heating and fan cooling. The entire system consumes a lot of power and is too bulky to be miniaturized. Handling of problems and requirements for rapid testing of samples;

[0007] 2. The volume of the photomultiplier tube or the charge-coupled element in the fluorescence detection system in the instrument is very large, and it is used separately, which requires a supporting optical path system, resulting in a large volume of the entire fluorescence micro-spectrum detection system, which increases the miniaturization

Cannot be applied in the rapidly developing space field;

[0008] 3. The transmission of excitation light and the collection of reflected light in the transmission system in the instrument require an optical path composed of various optical devices and optical fibers for optical transmission. Not only is the structure complex and difficult to miniaturize and integrate, but it also affects the sensitivity of each module in real-time fluorescence detection. and stability;

[0009] 4. The current instrument is not only bulky but also unable to monitor and feedback the temperature change in the microchannel, the flow rate of the microfluid and the fluorescence signal in real time;

[0010] 5. The medium temperature control system of small instruments lacks complete isolation, and the three temperature zones are easy to interfere with each other, which seriously affects the biological reaction results;

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