基于全光纤实时荧光定量核酸扩增技术的肿瘤相关基因高灵敏度检测方法研究

This project aims at shortening the detection period and improving the detection sensitivity for specific tumor-related genes based on real-time fluorescence quantitative Polymerase Chain Reaction (RT-PCR). The optical fiber microcavities used in this project demonstrate good biocompatibility and temperature sensing performance with low loss. By applying these properties and super-fast heating method with our laser system, we are able to explore the all-fiber RT-PCR for tumor-related genes sensing. In this project, a new fiber microcavity temperature circulating method based on laser super-fast heating system was proposed innovatively and we realized the RT-PCR in an all-fiber structure. By monitoring the fluorescence intensity, the sensing of specific tumor-related genes can be achieved with high efficiency, sensitivity and accuracy. It is even promising to realize single tumor-related gene sensing and improve the sensing accuracy dramatically of the existing technology. This proposed method is composed of optical, control science, chemistry and biomedical theory, and also can be potential utilized to detect genes related to Aids, thalassemia and hemophilia etc. This project will help improve the precaution of many kinds of severe illness and push the development of modern medicine.

本项目以缩短对特定肿瘤相关基因的检测时间、提高肿瘤相关基因的检测灵敏度为出发点，以实时荧光定量核酸扩增（PCR）技术为核心方法，结合光纤微腔良好的生物相容性、温度传感能力、低损耗特性以及激光超快温度加热方法，开展基于全光纤实时荧光定量PCR技术的肿瘤相关基因高灵敏度检测方法研究工作。本项目提出了一种基于激光超快加热系统的光纤微腔液体温度循环的新方法，拟在光纤中实现实时荧光定量PCR过程，并通过监测反应液荧光强度，实现对特定肿瘤相关基因的快速、高灵敏度检测，其检测精度可以达到单个肿瘤相关基因，大幅度提高目前肿瘤相关基因的检测灵敏度。这种光学、控制科学、化学、生物医疗的交叉融合技术将组成一套全新的测试手段，促进光纤传感在生物检测领域更好、更快地发展，同时该技术也可以拓展实现艾滋病、地中海贫血和血友病等重大传染病、遗传病相关基因的高灵敏度检测，大力推进重大疾病的防治以及现代医疗领域的跨越式发展。

本项目面向人民生命健康，以缩短对肿瘤相关基因的检测时间、简化检测步骤、降低检测成本为出发点，以光纤微腔技术、激光加热技术以及实时定量PCR技术为核心方法，以进一步推动肿瘤相关基因检测技术为研究目标，结合光纤微腔良好的生物相容性、可填充特性、温度传感能力、折射率传感能力、低损耗高聚焦特性以及激光系统超快温度加热方法，成功在光纤中实现免标记的实时定量PCR过程，通过监测反应液的荧光变化，实现了对特定肿瘤相关基因的快速、低成本、高灵敏度检测。结果表明，本系统能够成功实现ACTB内参基因以及肺癌基因EGFR的检测，检查灵敏度能够达到纳摩尔每升级别。在科研成果方面，以第一作者或通讯作者发表SCI检索论文13篇，其中在Sensors and Actuators B: Chemical和Analytical Chemistry等中科院1区期刊发表论文5篇，在IEEE Transactions on Instrumentation and Measurement，Journal of Lightwave Technology，IEEE Sensors Journal以及Optics Letters等中科院二区期刊发表论文8篇。本项目这种光学、控制科学、化学、生物医疗的交叉融合技术组成了一套全新的测试手段，促进光纤传感在生物检测领域更好、更快地发展。且本系统具有通用性，也可以拓展实现艾滋病、地中海贫血和血友病等重大传染病、遗传病相关基因的高灵敏度检测，推进重大疾病的防治以及现代医疗领域的发展。