一种分区温控的实时荧光PCR快速热循环系统设计
收稿日期: 2022-09-14
修回日期: 2022-11-14
录用日期: 2022-11-16
网络出版日期: 2023-09-27
Design of a Rapid Thermal Cycling System for Real-Time Fluorescent PCR with Zone Temperature Control
Received date: 2022-09-14
Revised date: 2022-11-14
Accepted date: 2022-11-16
Online published: 2023-09-27
热循环系统是实时荧光聚合酶链式反应(PCR)仪的关键组成部分,决定了核酸检测效率和结果准确性.针对传统PCR检测系统的热循环耗费时间长、温度控制复杂的问题,设计了一种分区温控的实时荧光PCR热循环系统,包括热循环系统硬件电路和机械结构,通过控制试液在不同恒温区切换实现快速热循环,采用增量式比例积分微分算法控制恒温区温度,控制精度达到 ±0.1 ℃.使用Fluent建立传热模型,分析试液热延迟现象预估试液变化规律.通过搭建PCR样机进行实验验证,试液升降温速率分别为3.8和4.4 ℃/s,证明了所提出PCR热循环系统能有效提高检测效率.
陈尔东, 高孜航, 王坤东, 雷华明 . 一种分区温控的实时荧光PCR快速热循环系统设计[J]. 上海交通大学学报, 2023 , 57(9) : 1196 -1202 . DOI: 10.16183/j.cnki.jsjtu.2022.361
The thermal cycling system is the key component of the real-time fluorescent polymerase chain reaction (PCR) instrument, which determines the nucleic acid detection efficiency and result accuracy. Aiming at the problems that the thermal cycle of the traditional PCR detection system takes a long time and the temperature control is complicated, a real-time fluorescent PCR thermal cycle system with partition temperature control is designed, including the hardware circuit and mechanical structure of the thermal cycle system. The rapid thermal cycling is achieved by switching the test solution between different constant temperature zones, and the temperature is controlled using an incremental proportional integral derivative (PID) algorithm, with a control precision of ±0.1 ℃. Using Fluent software to establish heat transfer model, the thermal delay phenomenon of the test solution was analyzed to predict the temperature variation pattern of the test solution. A prototype is built for testing and verification, and the heating and cooling rates of the test liquid are 3.8 ℃/s and 4.4 ℃/s. It is verified that the proposed PCR thermal cycling system can effectively improve the detection efficiency.
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