基于微流控芯片的荧光定量PCR法快速检测乙肝病毒核酸  

Rapid Detection of Hepatitis B Virus Nucleic Acid Based on Microfluidic Chip Using Fluorescence Quantitative PCR

作　　者：王可可 杨柯[1] 赵俊[1,2] 朱灿灿[1,2] 朱灵[1] 刘勇[1] WANG Ke-ke;YANG Ke;ZHAO Jun;ZHU Can-can;ZHU Ling.;LIU Yong(Institute of Applied Technology,Hefei Institutes of Physical Science,Chinese Academy of Sciences,Anhui Provincial Engineering Technology Research Center for Biomedical Optical InstrumentAnhui Provincial Engineering Laboratory for Medical Optical Diagnosis ~ Treatment Technology and Instrument,He f ei 230031;University of Science and Technology of China,Hefei 230026)

机构地区：[1]中国科学院合肥物质科学研究院应用技术研究所安徽省生物医学光学仪器工程技术研究中心安徽省医用光学诊疗技术与装备工程实验室,安徽合肥230031 [2]中国科学技术大学,安徽合肥230026

出　　处：《分析科学学报》2018年第4期450-454,共5页Journal of Analytical Science

基　　金：国家自然科学基金(No.61504152);安徽省科技重大专项(No.16030801123);安徽省自然科学基金(No.1608085QF135);安徽省国际科技合作计划(No.1704e1002224)

摘　　要：提出了一种基于微流控荧光定量聚合酶链式反应(Polymerase Chain Reaction,PCR)的血液病毒快速检测新技术。采用3D打印技术制作了一种集气动阀和"树型"结构为一体的微流控芯片;运用Comsol软件对芯片导热性能进行了仿真;根据血站血液核酸检测工作中乙型肝炎病毒(HBV)、丙型肝炎病毒(HCV)、人类免缺陷病毒(HIV)三种病毒的筛查流程,结合高温控性能的微流控荧光定量PCR分析系统,在所设计微流控芯片上开展了血液样本中HBV的检测研究。实验结果表明,该芯片具有良好的温度均匀性和导热性,芯片上可实现血液样本中HBV的快速检测,其Ct值在37左右呈弱阳性。该技术与目前血液筛查使用的大型核酸检测系统相比,具有操作过程简单、所占空间小、检测效率高且试剂消耗量少的优点,进一步优化实验条件可实现多病毒核酸的并行检测。A new technique for rapid and automated detection of blood virus based on microfluidic fluorescence quantitative polymerase chain reaction（PCR）was proposed.In this study,a microfluidic chip integrating with on-chip pneumatic valve and ＂tree＂ structure was fabricated by 3 D printing technology.The thermal conductivity of the chip was investigated by Comsol software.Based on the screening process of three conventional viruses HBV,HCV and HIV,the fluorescence quantitative PCR device with the high temperature control performance was combined for the HBV detection in blood samples.The experimental results showed that the microfluidic chip had excellent temperature uniformity and thermal conductivity.The rapid detection of HBV in blood samples can be realized on the designed microfluidic chip and the Ct value is about 37 showing weakly positive.Compared with the large-scale nucleic acid detection system used in blood center,our microfluidic chip has advantages of simple operations,small size,high detection efficiency and less reagent consumption,which can realize the parallel detection of multi-virus nucleic acid by further optimizing the experimental conditions.The results of the study have a good prospect in blood screening in the blood center and the field of point-ofcare testing such as preoperative blood screening.

关 键 词：3D打印 微流控芯片 HBV检测 Comsol 荧光定量PCR 

分 类 号：O657[理学—分析化学]