Research on the Key Technology of a Fluorescence Detection Device Using the RT-LAMP Method for Instant Detection

Abstract

As of 31 October 2023, there have been 771,795,258 confirmed cases of COVID-19 globally. Developing simple, portable, and reliable testing devices has become increasingly important. This paper presents a point-of-care testing (POCT) device for COVID-19 based on the dual-excitation fluorescence RT-LAMP method, which is derived from the principles of RT-LAMP-based COVID-19 detection kits available in the market. The key design solutions of the device were simulated and modeled. Key performance metrics such as detection repeatability and linearity were validated. Comparative experiments with the RT-qPCR detection method were conducted to verify the accuracy and reliability of the device. Additionally, the device’s detection sensitivity and accuracy were assessed. Experimental results show that the repeatability coefficient of variation (CV) value is ≤0.09%; the linearity R2 for the FAM channel is 0.9977 and that for the HEX channel is 0.9899; it exhibits good anti-interference performance, with negligible cross-channel interference; the temperature stability is ±0.062 °C, the temperature accuracy is less than 0.2 °C, and there is no significant temperature overshoot during the heating process. Compared with the real-time quantitative PCR (RT-qPCR) instrument, the positive agreement rate is 100% and the negative agreement rate is 95.0%. This research provides a foundational basis for the development of equipment for the prevention of infectious diseases and clinical diagnostics.