The combination of CRISPR-based biosensors with smartphone technology is changing point-of-care (PoC) diagnosis by offering fast, portable, and highly sensitive diagnosis of the disease. CRISPR/Cas biosensing has recently been further developed with Cas12a, Cas13a, and Cas14 proteins, and new signal transduction strategies, including DNAzymes, binary 3D DNA walkers, and photoelectrochemical sensing have been introduced to boost the sensitivity and specificity of CRISPR diagnostics for real-life use. Moreover, electrochemical and field-effect transistor (FET)-based biosensors, quantum dot-assisted fluorescence, and surface-enhanced Raman scattering (SERS) have also been used to enhance the sensitivity and accuracy of the signal.
Lab-on-a-Chip (LoC) microfluidic platforms have been integrated with diagnostic devices to achieve automated and multiplexed testing for high-throughput and low-cost testing strategies. AI-based smartphone applications now offer real-time data analysis, cloud-based result sharing, and machine learning-based interpretations of results to enhance diagnostic care and decision-making. In addition to nucleic acid detection, CRISPR systems engineered from bacteria and viruses have been deployed to recognize proteins, metabolites, and small molecules for their use in infectious disease surveillance, cancer screening, antimicrobial resistance detection, and environmental monitoring. However, there are several problems, including reagent stability, large scale clinical validation, and efficient signal amplification. A major concern with respect to CRISPR reagents is their robustness in different environmental conditions, and this remains an issue that requires urgent solutions.
The main challenges include developing isothermal amplification methods and obtaining regulatory signatures. In the future, research should be aimed at the application of graphene-based nanomaterials for the increase in the signal transfer rate, the optimization of the CRISPR reagents to have a long shelf life, and the creation of wearable biosensors for healthcare purposes. The combination of CRISPR technology, nanomaterials, artificial intelligence, and digital health solutions is likely to redefine decentralized diagnostics and extend globally to affordable and scalable healthcare solutions.
