Aptamers refer to a class of oligonucleotide probes that have demonstrated remarkable capabilities beyond mere genetic coding, including the unique ability to recognize and selectively bind to specific molecular targets. Numerous advantages, including accessibility for targeting a diverse array of molecules and compatibility with different signal amplification and transduction elements, underscore the application of aptamers for delivering rapid and accurate diagnostic tests at the point of care. This review provides a comprehensive summary of the recent advances in aptamer-based point-of-care testing, especially highlighting the innovative applications of aptamers in colorimetric sensors, lateral flow assays, fluorescent biosensors, and electrochemical biosensors. Additionally, current challenges in this burgeoning field and forward-looking perspectives for aptamer-based point-of-care testing are discussed.