Real-time Fluorescent Quantitative PCR Device Integrated With Sample Pretreatment

Real-time fluorescent quantitative PCR has always been the mainstream method to detect the nucleic acid concentration of diseases.Traditional laboratory or hospital instruments need to go through the tedious nucleic acid pretreatment process,and then amplify the extracted nucleic acid.Usually,this kind of instrument is accompanied by high cost,long time-consuming,low integration and professional operation.In this paper,a real-time fluorescence quantitative PCR device integrated with sample pretreatment was designed for the scene of on-site detection,and the related technologies were studied.The work of this paper is summarized as follows:1.A porous polydimethylsiloxane(PDMS)chip was designed.Combined with a micro slide table to control the movement of the permanent magnet,the extraction of nucleic acid by magnetic beads was realized.The whole process was completed in 15 minutes.2.The whole instrument size is 45 cm*25 cm*32 cm,and the power supply of the system is 12 V,which is convenient for on-site detection.The upper computer software suitable for image processing and data analysis of the system is developed.3.Compared with ABI7900 real-time fluorescence quantitative PCR instrument,the developed real-time fluorescence quantitative PCR instrument used the same concentration of samples to do multiple groups of repeated experiments,which showed that the instrument had good linearity,and the detection line was 100 copies/μL.4.Using Mask R-CNN method to explore the problem of fluorescent PCR image recognition,the accuracy and recognition rate are improved compared with the traditional threshold segmentation method.The system developed in this paper has the characteristics of high integration,low cost and low power consumption,which is suitable for the application of field detection.