Research And Realization Of Key Technologies For PCR Amplification And Detection System

PCR(Polymerase Chain Reaction)amplification technology is an important detection method for various diseases,and is even regarded as one of the gold standards for the detection of infectious diseases.The advantages of PCR nucleic acid detection include:high sensitivity,strong specificity,high accuracy,and high technology maturity.The traditional PCR amplification reaction needs to be supported by a thermal cycler that integrates a sophisticated precision temperature control system.Therefore,the PCR instrument is large in size,high in cost,and complicated in operation.It has high requirements on laboratory hardware conditions and operators,which is not conducive to promotion and application of PCR amplification detection technology.In addition,the traditional PCR instrument uses a test tube as a reaction carrier.Due to the influence of the heat capacity of heating module and test tube as well as the thermal inertia of the entire heating link,thermal cycling processes tend to have a lower temperature rise and fall rate,resulting in the switching process between each reaction temperature takes up more total PCR reaction time,thus making the PCR total detection time gets longer.In order to further improve the comprehensive performance of PCR detection,it is of great significance to combine microfluidic technology to develop fast,simple,low-cost and convenient PCR detection technologies and systems for the POCT(Point-of-Care Testing)and the realization of nucleic acid detection in various medical environments,atypical medical environments,community hospitals,clinics,and even outdoors.This paper studies the key technologies related to PCR amplification and detection.On the one hand,combined with the microfluidic chip technology,the PCR amplification technology and system based on the microfluidic chip were studied to realize portable,automated and rapid PCR amplification.The experimental results show that the microfluidic PCR amplification system can achieve successful amplification detection of amplified samples.On the other hand,based on the principle of CPCR(Convective PCR)amplification and fluorescence detection,high-throughput real-time CPCR amplification technology and system were studied.Compared with the traditional PCR amplification method(amplification time:1-2 hours),the high-throughput real-time CPCR amplification system based on simplified constant temperature heating mode,which not only significantly reduces the cost of the instrument,but also significantly shortens the nucleic acid detection time(within 30 minutes)and achieves ultra-high speed nucleic acid amplification detection.The experimental results show that the high-throughput real-time CPCR amplification system can quickly realize the amplification and detection of specific targets.Based on the obvious advantages in terms of speed,convenience,size,cost and portability,the microfluidic PCR amplification system and high-throughput real-time CPCR amplification system laid a solid foundation for POCT nucleic acid detection.