连续流PCR芯片DNA扩增技术的研究  

Amplification of DNA based on continuous flow PCR microfluidic chips

作　　者：程抒一 陶春先[1] 庄松林[1] 张大伟[1] 李振庆[1] 山口佳则[2] CHENG Shu-yi;TAO Chun-xian;ZHUANG Song-lin;ZHANG Da-wei;LI Zhen-qing;YAMAGUCHI Yoshinori(School of Optical-Electrical and Computer Engineering,Key Laboratory of Medical Optical Instrument and Equipment of Ministry of Education,University of Shanghai for Science and Technology,Shanghai 200093;Photonics Advanced Research Center,Graduate School of Engineering,Osaka University,Yamadaoka Suita-city,Osaka 565-0871,Japan)

机构地区：[1]上海理工大学光电信息与计算机工程学院,教育部医用光学仪器与设备重点实验室,上海200093 [2]大阪大学先进光子研究中心,大阪565-0871

出　　处：《分析试验室》2018年第12期1416-1420,共5页Chinese Journal of Analysis Laboratory

基　　金：上海市“科技创新行动计划”生物医药领域科技支撑项目(18441900400)资助

摘　　要：提出了一种基于二温区的连续流PCR微流控芯片,并通过有限元方法分析了其内部流速场与温度场分布,数值分析结果显示:(1)芯片中液体流速高于5.128mm/s时,液体在微通道中心流速与靠近内壁的流速差异较大,不利于PCR反应;(2)当CF-PCR微流控芯片两侧施加96℃和65℃高温时,液态水可以在0.1mm内迅速实现变温,且中间未施温部分保持在73℃,为DNA实现PCR提供了变性、退火与延伸所需温度。实验结果表明,压力法进样可使流速误差稳定在3%以内,采用铝块对PCR芯片加热可使温度误差控制在0.3%。最后在CF-PCR芯片内对牙龈卟啉单胞菌实施了PCR,并用毛细管电泳检测了其PCR产物,数据显示在芯片内4′51″可以成功实现PCR。本方法对快速PCR仪器的开发具有重要的应用价值。A two temperature zone continuous flow PCR( CF-PCR) microfluidic chip was proposed and the flow distribution and temperature in the CF-PCR chip was analyzed by finite element analysis.Results demonstrated that when the velocity was above 5. 128 mm/s,the velocity of the liquid in the center of the micro-channel was quite different from the one from the inner wall,which was adverse to the PCR. When applying 96℃ and 65℃ on both side of the CF-PCR chip,the temperature of water can be quickly changed within 0. 1 mm,and the middle part of the chip kept at 73℃. Therefore,twotemperature zone provided the temperature for denature,annealing,and extension of DNA,which was needed for PCR. Experiments showed that injection of the sample by pressure could make the velocity error less than 3%. Heating the chip by aluminum could make the temperature error less that 0. 3%.The PCR of P. g in the chip was performed and the PCR products was analyzed by capillary electrophoresis. Results showed that PCR was successfully realized within 4’ 51″. This method had great value for the development of fast PCR instrument.

关 键 词：微流控芯片 聚合酶链式反应 毛细管电泳 

分 类 号：TS83[轻工技术与工程] O657.8[理学—分析化学]