Molecular assays and point-of-care tools: a review of amplification methods and nucleic acid detection systems

Molecular diagnosis is crucial for combating infectious diseases due to its high sensitivity and precision. However, its application in point-of-care testing (POCT) faces challenges, particularly in remote areas and developing countries. This review, based on searches of indexed publication databases, examined major nucleic acid amplification methods, detection techniques and miniaturization strategies for POCT platform development, focusing on the period 2015–2025. The need for regulatory standardization and the advantages and disadvantages of amplification techniques were discussed. Microfluidic approaches—reaction chambers, continuous flow, and droplets—were analyzed as the basis for functional device integration, emphasizing their role in reducing volumes, energy consumption and processing time. The sensitivity, cost and instrumentation complexity of fluorescence-, colorimetry- and electrochemistry-based detection systems were addressed. The practical feasibility of integrating these into miniaturized POCT platforms underscores the structural role of microfluidics in advancing portable molecular diagnostics. Despite progress, miniaturization complexity and reagent costs remain significant technological barriers.

Key words
Amplification; Infectious Diseases; Molecular Diagnostics; Nucleic Acid; Pathogen Detection; Point-of-care Testing