Abstarct:

The advent of CRISPR-associated (Cas) systems has dramatically transformed molecular diagnostics, enabling unmatched accuracy, speed, and portability for detecting pathogens. Since their first use in 2016, CRISPR-based diagnostic tools have progressed from experimental concepts to clinically validated systems capable of identifying viral, bacterial, and genetic targets with attomolar sensitivity. This thorough review explores the current landscape of CRISPR molecular diagnostics, emphasizing significant technological platforms such as SHERLOCK, DETECTR, and new emerging systems. We explore the fundamental principles of Class II CRISPR-Cas systems (Cas9, Cas12, and Cas13), their synergy with isothermal amplification methods, and their groundbreaking applications in detecting infectious diseases, especially in relation to COVID-19 and other emerging pathogens. The review highlights essential challenges like sample processing limitations, stability issues, multiplexing abilities, and regulatory obstacles that currently hinder widespread clinical use. Moreover, we examine recent advancements in point-of-care diagnostics, the integration of artificial intelligence, and the progress in developing amplification-free detection systems. Finally, we offer insights into future prospects for CRISPR-based molecular diagnostics, including their potential impact on personalized medicine and global health in resource-constrained environments.