Recent advances in isothermal amplification techniques coupled with clustered regularly interspaced short palindromic repeat/Cas systems

Abstract

Isothermal amplification (IA) techniques have emerged as promising alternatives to polymerase chain reaction (PCR), enabling rapid and efficient nucleic acid amplification at constant temperatures. Meanwhile, clustered regularly interspaced short palindromic repeat (CRISPR)/Cas‐based technology has revolutionized molecular diagnostics by harnessing collateral cleavage for programmable biomarker detection. Especially, the combination of IA techniques with CRISPR/Cas systems establishes a synergistic framework for next‐generation diagnostic platforms, addressing the sensitivity‐specificity trade‐offs and challenges posed by resource‐limited settings inherent in conventional methods. In this review, we trace the developmental milestones of IA techniques and elucidate the fundamental mechanisms of relevant IA techniques and CRISPR/Cas systems. Synergistic strategies for coupling IA techniques with CRISPR/Cas systems were summarized. Subsequently, personalized applications including point‐of‐care testing devices, droplet microfluidics platforms for rare biomarker detection, and IA/CRISPR/Cas‐based nanosensors were introduced. Finally, several challenges and perspectives in the field are discussed.