Handheld Continuous-flow Real-time Fluorescence qPCR System with PVC Microreactor
The polymerase chain reaction (PCR) has unique advantages of sensitivity, specificity and rapidity in pathogen detection, which makes it at the forefront of academia and application in molecular biology diagnosis. In this study, we proposed a hand-held real-time fluorescence qPCR system, which can be used for the quantitative analysis of nucleic acid molecules. For the first time, we use a PVC microreactor which improved the transmittance of the microreactor and the ease to collect the fluorescence signal. In order to make it portable, the system adopted the passive syringe for sample injection and integrated temperature control and detection with a lithium battery for power supply. What’s more, the fluorescence signal was captured by a smartphone through an external automatic robotic arm. This real-time qPCR system can detect genomic DNA of the H7N9 avian influenza with over four orders of magnitude concentration from 107 to 104 copies per μL. In addition, it verified that the fluorescence images obtained by this system were clearer than those obtained by traditional system (using PTFE spatial PCR microreactor) with two typical dyes and a probe tested——EvaGreen, SYBR Green and FAM.