Research On Single-Microchannel Based Fluorescence PCR System And Its Key Technology

With steady pace of China's space dream “Three Step Walk” strategy, China has achieved multiple technical accomplishments including multi-manned and multi-day space flight, spacecraft and space cabin connecting, which represent a new era of national space exploration. Due to the special space environment, it is crucial to ensure the health and safety of researchers by having long-term real-time environment monitoring at space station. Therefore, it is imperative to build a space-applicable real-time automatic microbiological hazard alarm system.This paper proposed four, with independent intellectual property, integrated single-microchannel real-time fluorescence PCR systems, which represents the core technology of real-time automatic microbiological hazard alarm system, and carried the following research.(1) Analysis on four single-microchannel based fluorescence PCR systems is conducted and reciprocating circular single-microchannel system has shown simplified system structure, direct, stable, accurate and highly integrated power source,which has typical research significance. Compared to traditional 40 circular porous multi-row design, single-microchannel design has faster response rate, reduce reagent retention, lower overall system consumption, higher detection pertinence and higher overall response efficiency.(2) It is of great significance, for space micro-volume high-sensitivity optical design, to build the micro-fluorescence detection system of single-microchannel fluorescence PCR system and make integrated design of stable LED light source and1 mm x 1mm micro-size detection sensor. After experiments, In this fluorescence detection module test, the relative value of the negative reagent is about 0.1V, and the relative value of the positive reagent is about 0.33 V. The fluorescence relative value of this system is more than 3 times, and it is satisfied with the original basis of the quantitative analysis curve of the results of PCR amplification reaction.(3) Temperature control is done through the coordination of environment field detection and temperature zone point detection of the single-microchannel fluorescence PCR system. Environment field detection measures the value range, and temperature zone point detection measures the exact value. The overall system consists of circuit designs of multi-sensor signal acquisition, LCD display and keyboard input. PID algorithm is used for temperature zone real-time detection and closed-loop control heating to ensure a constant temperature. The detection results of single-microchannel fluorescence PCR system show that PCR amplification reaction zone has constant temperature, the temperature control precision is 1.5 ?, and the average signal acquisition error is around 1.0?.