用于微流控荧光PCR的高分辨率温度控制系统

大气与环境光学学报 ›› 2015, Vol. 10 ›› Issue (5): 417-424.

用于微流控荧光PCR的高分辨率温度控制系统

范书广1, 朱灵1,2*, 赵树弥1, 张悦龙1, 张龙1,2, 邓国庆1,2, 王安1, 刘勇1,2

（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，安徽合肥 230031;
2 皖江新兴产业技术发展中心，安徽铜陵244000）

收稿日期:2014-11-11 修回日期:2014-12-04 出版日期:2015-09-28 发布日期:2015-09-11
作者简介:范书广（1989-），男，安徽阜阳人，研究生，主要研究方向为光学精密仪器的研发。
基金资助:
中国科学院战略性先导科技专项基金项目（XDA08040109）资助

High Resolution Temperature Control System of Microfluidic Fluorescence PCR

FAN Shuguang1, ZHU Ling1,2*, ZHAO Shumi1, ZHANG Yuelong1, ZHANG Long1,2, DENG Guoqing1,2, WANG An1, LIU Yong1,2*

(1 Anhui Provincial Engineering Technology Research Center for Biomedical Optical Instrument, Institute of Applied Technology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, China;
2 Wanjiang Center for Development of Emerging Industrial Technology, Tongling 244000, China)

Received:2014-11-11 Revised:2014-12-04 Published:2015-09-28 Online:2015-09-11
Contact: 范书广（1989-），男，安徽阜阳人，研究生，主要研究方向为光学精密仪器的研发。 E-mail:zhul@aiofm.ac.cn

摘要/Abstract

摘要：

微流控荧光PCR产物在高分辨率熔解（HRM）曲线分析的过程中，随着DNA的解链荧光强度发生突变，而能否检测到该突变点则依赖于温控的精度。提出了多阈值PID算法，设计了一种适用于高分辨率熔解曲线分析的FPGA温度控制系统。根据测得温度值与设定值之间的差异，改变FPGA电路系统输出的脉冲宽度调制(pulse width modulation, PWM)波占空比和算法的调节频率实现对温度的准确控制。经过温度标定后，系统温度控制精度优于0.1℃，分辨率达到±0.01℃。HRM分析实验结果表明，本方法成功地辨别出了DNA序列的单碱基差异，温度控制系统达到了设计的要求。

关键词: 微流控, 铂电阻, 多阈值PID算法, 脉冲宽度调制波, 高分辨率熔解

Abstract:

In the analysis of high resolution melting (HRM) of microfluidic fluorescence PCR amplification products, fluorescence intensity has a sudden change with the delink of DNA chain, while whether the abrupt change can be detected is dependent on the accuracy of temperature control. A kind of multi threshold PID algorithm was put forward, and a temperature control system based on FPGA was designed to adapt to the analysis of high resolution melting. Based on the difference of the measurement value and the set value, the temperature change can be controlled accurately through changing the pulse-time ratio of pulse width modulation(PWM) wave and the control frequency of the algorithm from the FPGA circuit system. After temperature calibrations of the measurement system, the accuracy precedes 0.1℃ and the resolution achieves ±0.01℃. The result of HRM experiment shows that this system can detect the single base mutation of DNA sequence successfully, and the temperature control system can meet the design requirements

Key words: microfluidic, platinum resistance, multi threshold PID algorithm, pulse width modulation wave, high resolution melting

中图分类号:

R318.6

范书广朱灵赵树弥张悦龙张龙邓国庆王安刘勇. 用于微流控荧光PCR的高分辨率温度控制系统[J]. 大气与环境光学学报, 2015, 10(5): 417-424.

FAN Shu-Guang, ZHU Ling, ZHAO Shu-Mi, Zhang Yue-Long, ZHANG Long, DENG Guo-Qing, WANG An, LIU Yong. High Resolution Temperature Control System of Microfluidic Fluorescence PCR[J]. Journal of Atmospheric and Environmental Optics, 2015, 10(5): 417-424.

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