Comprehensive Review on Viral RNA Extraction Strategies for Enhanced Molecular Diagnostics

Abstract

RNA viruses, characterized by their RNA-based genomes, have garnered significant concern due to their role in numerous lethal pandemics and remain a significant global health threat. Their predominance among emerging pathogens underscores the need for rapid detection and molecular characterization. PCR has been proven to be an indispensable tool for monitoring the emergence and spread of RNA viruses, but its success requires efficient viral RNA isolation. Poor RNA recovery may lead to failed detection, limiting downstream applications such as genome sequencing and molecular epidemiology. This review provides an overview of major RNA extraction strategies, including TRIzol-based, spin column–based, and magnetic bead–based methods, as well as novel and automated platforms. We compared their advantages, limitations, and trade-offs, emphasizing compatibility with downstream processes such as PCR and next-generation sequencing. Challenges related to sample collection, transport, storage, and RNA integrity are discussed, particularly in resource-limited settings where cold-chain and biosafety infrastructure may be limited. Additionally, we discussed applications of extracted RNA for diagnostics, viral characterization, and public health interventions, as well as advances in point-of-care (POC) technologies that integrate extraction with detection. Future directions focus on developing robust, cost-effective, and scalable extraction methods that are adaptable to both high-throughput laboratories and portable POC systems, ensuring global preparedness against emerging RNA viruses.