Recent Advances in Microfluidic-Based Biosensors for Nucleic Acid Detection: From Amplification Strategies to...

Nucleic acid detection has gained significant attention in healthcare because of its crucial role in diagnosing infectious and non-infectious diseases. Conventional molecular diagnostics are predominantly performed in centralized laboratories using sophisticated instrumentation and automated analyzers, resulting in increased analysis time, cost, and limited accessibility in decentralized settings. Consequently, there is an increasing demand for rapid, cost-effective, portable, and user-friendly diagnostic platforms capable of providing reliable results at the point of care. Target nucleic acid detection commonly relies on amplification strategies, including thermocyclic amplification such as polymerase chain reaction (PCR) and various isothermal amplification techniques. Although these methods provide high sensitivity and specificity, challenges associated with instrumentation, thermal control, sample preparation, assay integration, and accessibility remain. Microfluidic integration provides a promising approach for addressing these limitations by enabling miniaturized sample handling, reduced reagent consumption, rapid amplification, and integrated detection within compact analytical platforms. This review critically examines recent advances in microfluidic nucleic acid biosensors, including PCR and isothermal amplification platforms, emerging amplification and detection technologies, device integration, and point-of-care applications. Further, emphasis on comparative analytical performance, technological limitations, and the remaining challenges associated with translating microfluidic nucleic acid detection systems from laboratory demonstrations to practical point-of-care diagnostics applications.