Development of Nucleic Acid Isothermal Amplification Technologies for Virus Detection

Viruses play a significant role in causing human diseases, and traditional PCR techniques have been widely used for their molecular diagnosis. However, the temperature requirements of PCR limit its application in field diagnostics. To address the need for rapid on-site diagnosis, isothermal nucleic acid amplification technologies have emerged as a promising alternative. These technologies enable nucleic acid amplification at a constant temperature without the need for thermal cycling, making them more adaptable for different diagnostic settings. This comprehensive review examines the latest advancements in isothermal amplification technologies for virus detection. It covers various aspects, including viral sample collection, nucleic acid extraction, and isothermal amplification detection. The review explores the principles, key parameters, and applications of enzyme-assisted isothermal amplification, enzyme-free isothermal amplification, and cascade amplification techniques integrated with multiple systems. Furthermore, a comparison of commercially available reagent kits is provided to highlight their respective characteristics. Additionally, the review discusses the current challenges faced by isothermal nucleic acid amplification technologies in pathogen detection, such as extraction efficiency, stability, and cost, and proposes future directions to enhance the on-site diagnostic efficacy of these technologies.