Abstract
Alpha-fetoprotein (AFP), commonly employed for early diagnosis of liver cancer, serves as a biomarker for cancer screening and diagnosis. Combining the high sensitivity and specificity of fluorescence immunoassay (FIA), developing a low-cost and efficient immunoassay system for AFP detection holds significant importance in disease diagnosis. In this work, we developed a miniaturized oblique laser-induced fluorescence (LIF) immunoassay system, coupled with a microfluidic PMMA/paper hybrid chip, for rapid detection of AFP. The system employed an avalanche photodiode (APD) as the detector, and implemented multi-level filtering in the excitation light channel using the dichroic mirror and optical trap. At first, we employed the Savitzky–Golay filter and baseline off-set elimination methods to denoise and normalize the original data. Then the cutoff frequency of the low-pass filter and the reverse voltage of the APD were optimized to enhance the detection sensitivity of the system. Furthermore, the effect of laser power on the fluorescence excitation efficiency was investigated, and the sampling time during the scanning process was optimized. Finally, a four-parameter logistic (4PL) model was utilized to establish the concentration–response equation for AFP. The system was capable of detecting concentrations of AFP standard solution within the range of 1–500 ng/mL, with a detection limit of 0.8 ng/mL. The entire immunoassay process could be completed within 15 min. It has an excellent potential for applications in low-cost portable diagnostic instruments for the rapid detection of biomarkers.
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The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
This research was supported by the National Natural Science Foundation of China (Grant No. 62304057), Guangxi Bagui Scholars Project (No.2019A02), Guangxi University Innovation Development Doubling Plan project (No.202201343 and No.202201369),and Science and Technology Major Project of Guangxi (Guike AA22117005 and Guike AA22117007).
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Zheng, C., Dai, P., You, H. et al. A compact microfluidic laser-induced fluorescence immunoassay system using avalanche photodiode for rapid detection of alpha-fetoprotein. ANAL. SCI. (2024). https://doi.org/10.1007/s44211-024-00553-3
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DOI: https://doi.org/10.1007/s44211-024-00553-3