Validation of improved automated nucleic acid extraction methods for direct detection of polioviruses for global polio eradication

J Virol Methods. 2024 Mar 6:114914. doi: 10.1016/j.jviromet.2024.114914. Online ahead of print.ABSTRACTPolioviruses (PV), the main causative agent of acute flaccid paralysis (AFP), are positive-sense single-stranded RNA viruses of the family Picornaviridae. As we approach polio eradication, accurate and timely detection of poliovirus in stool from AFP cases becomes vital to success for the eradication efforts. Direct detection of PV from clinical diagnostic samples using nucleic acid (NA) extraction and real-time reverse transcriptase polymerase chain reaction (rRT-PCR) instead of the current standard method of virus isolation in culture, eliminates the long turn-around time to diagnosis and the need for high viral titer amplification in laboratories. An essential component of direct detection of PV from AFP surveillance samples is the efficient extraction of NA. Potential supply chain issues and lack of vendor presence in certain areas of the world necessitates the validation of multiple NA extraction methods. Using retrospective PV-positive surveillance samples (n=104), two extraction kits were compared to the previously validated Zymo Research Quick-RNATM Viral Kit. The Roche High Pure Viral RNA Kit, a column-based manual extraction method, and the Magmax Pathogen RNA/DNA kit used in the automated Kingfisher Flex system were both non-inferior to the Zymo kit, with similar rates of PV detection in pivotal rRT-PCR assays, such as pan-poliovirus (PanPV), poliovirus serotype 2...

Source: Journal of Virological Methods - March 8, 2024 Category: Virology Authors: Stacey Jeffries Miles Chelsea Harrington Hong Sun Ashley Deas M Steve Oberste W Allan Nix Everardo Vega Nancy Gerloff Source Type: research