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.

Waterborne pathogens seriously threaten human life and can cause diarrhea, gastrointestinal disorders, and more serious systemic infections. These pathogens are usually caused by contaminated water sources that contain disease-causing microorganisms, such as bacteria, viruses, and parasites, which cause infection and disease when they enter the human body through drinking water or other means. Due to the wide range of transmission routes and the high potential risk of waterborne pathogens, there is an urgent need for an ultrasensitive, rapid, and specific pathogenic microorganism monitoring platform to meet the critical monitoring needs of some water bodies’ collection points daily monitoring needs. Microfluidics-based pathogen surveillance methods are an important stage towards automated detection through real-time and multi-targeted monitoring, thus enabling a comprehensive assessment of the risk of exposure to waterborne pathogens and even emerging microbial contaminants, and thus better protection of public health. Therefore, this paper reviews the latest research results on the isolation and detection of waterborne pathogens based on microfluidic methods. First, we introduce the traditional methods for isolation and detection of pathogens. Then, we compare some existing microfluidic pathogen isolation and detection methods and finally look forward to some future research directions and applications of microfluidic technology in waterborne pathogens monitoring.

Tetrodotoxin (TTX) is a potent marine neurotoxin, necessitating sensitive and user-friendly on-site assays. To address long workflows of traditional immunoassays and limited signal amplification in colorimetric microfluidics, we developed a nanozyme-catalyzed colorimetric magnetic microfluidic immunosensor (Nano-CMI). This platform combines an aptamer–antibody sandwich capture format with catalytic amplification via AuNR@Pt@m-SiO₂ (APMS) nanozymes on a magnetically actuated microfluidic chip. Magnetic actuation simplifies sample handling and washing, while APMS catalysis enhances sensitivity and visual readout. The Nano-CMI has been used for the detection of TTX samples ranging from 0.2 to 20 ng/mL with a detection limit of 0.2 ng/mL in 10 min, following the linear equation: y = −31.14ln x + 110.15, and the entire “capture-reaction-detection” workflow can be completed within 1 h. With rapid response, minimal hands-on time, and robust performance, this platform offers a practical, high-sensitivity solution for on-site TTX screening in food safety and customs inspection.