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 made it easy to collect the fluorescence signal. In order to make it portable, the system adopted a 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 using a smartphone through an external automatic robotic arm. This real-time qPCR system can detect genomic DNA of the H7N9 avian influenza over four orders of magnitude of concentration from 10⁷ to 10⁴ copies per μL. In addition, it was verified that the fluorescence images obtained by this system were clearer than those obtained by a traditional system (using a PTFE spatial PCR microreactor) with two typical dyes and a probe tested—EvaGreen, SYBR Green and FAM.

中文翻译：

---

带有PVC微反应器的手持式连续流实时荧光定量PCR系统

聚合酶链反应（PCR）在病原体检测方面具有灵敏度，特异性和快速性的独特优势，使其在学术界和分子生物学诊断中具有领先地位。在这项研究中，我们提出了一种手持式实时荧光定量PCR系统，可用于核酸分子的定量分析。我们首次使用了PVC微反应器，该反应器提高了微反应器的透射率，并使其易于收集荧光信号。为了使其便携，该系统采用无源注射器进行样品注入，并集成温度控制和检测功能以及锂电池供电。此外，荧光信号是通过使用智能手机通过外部自动机械臂捕获的。每微升⁷至10 ⁴份。此外，已证实该系统获得的荧光图像比传统系统（使用PTFE空间PCR微反应器）使用两种典型染料和经过测试的探针-EvaGreen，SYBR Green和FAM获得的荧光图像更清晰。