Helicase-dependent isothermal amplification turns biosensing into modern diagnostic technologies

Helicase-dependent amplification (HDA) is an isothermal DNA amplification technique that utilizes the enzymatic activity of helicases to unwind double-stranded DNA, enabling the amplification of specific target sequences without the need for temperature cycling. This review provides an overview of the principles of HDA, focusing on the role of DNA helicase families and how they contribute to the efficient and specific amplification of nucleic acids. Furthermore, we analyze the diverse applications of HDA in point-of-care (POC), highlighting its potential to revolutionize diagnostic technologies by enabling rapid, portable, and cost-effective detection of pathogens, genetic mutations, and biomarkers. Additionally, other applications of HDA in diagnostics, such as detection of environmental contaminants and foodborne pathogens, are discussed. A comparison of HDA with conventional PCR and other isothermal amplification techniques reveals several advantages, including its simplicity, speed, and minimal equipment requirements, though challenges related to sensitivity, specificity, and scalability remain. Therefore, HDA emerges as a promising tool for advancing modern diagnostic technologies, particularly in resource-limited settings.