Point-of-care testing (POCT) is an innovative diagnostic technology that provides cost-effective and rapid analysis, as well as accurate diagnostics. It enables patients to obtain clinically relevant results through self-testing. This technology has played a vital role in clinical diagnosis, disease monitoring, and early detection of infectious diseases. Nucleic acid aptamers, which are molecular probes capable of specifically recognizing multiple targets, have emerged as valuable components in biomedical sensors for molecular recognition. They offer advantages such as easy synthesis, good stability, and signal amplification. In recent years, research on aptamer-based POCT technology has garnered widespread attention in the world. The key issues in current research include obtaining more high-affinity aptamers to meet the detection needs of various targets, improving detection sensitivity through signal amplification, and integrating with different sensors to meet the requirements of POCT products. In this review, we first briefly introduce the selection process and the targets used for systematic evolution of ligands by exponential enrichment (SELEX). We discuss new SELEX strategies that have been developed to improve the selection efficiency and enhance the affinity of aptamers. Furthermore, we analyze 4 commonly used signal amplification strategies in aptamer-based POCT sensors. Among these methods, nucleic acid signal amplification and self-assembly signal amplification techniques are commonly used due to their low cost and wide applicability. The combination of these two techniques has also been developed to improve detection sensitivity and shorten reaction time. Coupling aptamers with enzyme-based reactions is the simplest method to improve signal amplification in POCT sensors. Various nanomaterials, such as metal nanoparticles, graphene, carbon nanotubes, and metal-organic frameworks, are widely used to improve the detection sensitivity. The combination of multi-functional nanomaterials for signal amplification has also been introduced in this part. Additionally, we introduce a strategy that involves the use of aptamers to initiate the activation of CRISPR-associated proteins, resulting in the cleavage of DNA or RNA molecular beacons and leading to signal amplification. Furthermore, we also introduce the most recent advances in the development of various aptamer-based electrochemical sensors and optical sensors in the field of POCT. Aptamer-based electrochemical sensors offer advantages such as fast response, easy operation, and portability. In this part, we highlight a series of blood glucose meter based aptasensors used to quantify a variety of biomarkers. For the importance of research on continuous detection device, we review recent progress in the development of aptamer-based continues electrochemical testing devices. In aptamer-based optical POCT techniques, the recent development of colorimetry, lateral flow assay (LFA), fluorescence, surface-enhanced Raman scattering (SERS), surface plasmon resonance (SPR), and evanescent wave fiber optic sensors are introduced, with a focus on comparing the performance characteristics of each sensor. Finally, this review presents a summary and future challenges in the research and commercialization of aptamer-based POCT sensors. To simplify the aptamers selection process, it is crucial to invest in studying the molecular recognition mechanisms of aptamers and developing artificial intelligence-based methods for aptamer selection. Additionally, integrating aptamers with advanced sensor technologies like microfluidic chips and wearable devices can greatly enhance detection sensitivity and stability. From a commercial perspective, current aptamer-based POCT products mostly comprise fluorescent or colorimetric assay kits and lateral flow test strips. However, to garner more attention in the competitive POCT market, aptamer-based POCT sensors have an edge in small molecules detection and multi-channel detection.