基于超大型半导体致冷片快速升降温加热模块的核酸快速扩增技术的实现

Biosensors Pub Date : 2024-08-06 DOI:10.3390/bios14080379
Jianxin Cheng, Enjia Zhang, Rui Sun, Kaihuan Zhang, Fangzhou Zhang, Jianlong Zhao, Shilun Feng, Bo Liu
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引用次数: 0

摘要

在分子生物学飞速发展的今天,核酸扩增检测技术越来越受到重视。传统的聚合酶链反应(PCR)仪器在温度循环中从高温向低温过渡时制冷性能较差,导致 PCR 扩增周期较长。因此,我们采用了同时具备加热和冷却功能的珀尔帖元件,并利用稳健的自适应模糊比例积分导数(PID)算法作为基本的温度控制机制。加热和冷却功能通过状态机模式进行切换,PCR 温度控制模块的设计旨在实现快速的温度变化。循环温度测试结果表明,采用模糊 PID 控制算法可以准确控制温度,实现快速升降温(平均升温速度 = 11 ℃/s,平均降温速度 = 8 ℃/s),同时防止温度过高,保持温度稳定,实现超快速 PCR 扩增过程(45 个温度循环时间小于 19 分钟)。定量结果表明,根据添加的病毒颗粒浓度不同,可观察到不同数量的荧光信号,分析检测限(LoD)低至每微升 10 个拷贝,阴性对照无假阳性。结果表明,TEC 核酸扩增法具有较高的检测率、灵敏度和稳定性。这项研究旨在解决现有热循环温度控制技术难以满足各种新的发展要求的问题,如 PCR 的快速、高效和微型化。
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Implementation of Rapid Nucleic Acid Amplification Based on the Super Large Thermoelectric Cooler Rapid Temperature Rise and Fall Heating Module
In the rapid development of molecular biology, nucleic acid amplification detection technology has received more and more attention. The traditional polymerase chain reaction (PCR) instrument has poor refrigeration performance during its transition from a high temperature to a low temperature in the temperature cycle, resulting in a longer PCR amplification cycle. Peltier element equipped with both heating and cooling functions was used, while the robust adaptive fuzzy proportional integral derivative (PID) algorithm was also utilized as the fundamental temperature control mechanism. The heating and cooling functions were switched through the state machine mode, and the PCR temperature control module was designed to achieve rapid temperature change. Cycle temperature test results showed that the fuzzy PID control algorithm was used to accurately control the temperature and achieve rapid temperature rise and fall (average rising speed = 11 °C/s, average falling speed = 8 °C/s) while preventing temperature overcharging, maintaining temperature stability, and achieving ultra-fast PCR amplification processes (45 temperature cycle time < 19 min). The quantitative results show that different amounts of fluorescence signals can be observed according to the different concentrations of added viral particles, and an analytical detection limit (LoD) as low as 10 copies per μL can be achieved with no false positive in the negative control. The results show that the TEC amplification of nucleic acid has a high detection rate, sensitivity, and stability. This study intended to solve the problem where the existing thermal cycle temperature control technology finds it difficult to meet various new development requirements, such as the rapid, efficient, and miniaturization of PCR.
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