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

• DOI: 10.1016/j.jviromet.2024.114914
• Publication Date: 2024-03-06T17:16:30Z
• Authors: Stacey Jeffries M...; Chelsea Harrington; Hong Sun; Ashley Deas; M. Steven Oberste; W. Allan Nix; Everardo Vega; Nancy Gerloff
• Keywords: Molecular diagnostics

Polioviruses (PV), the main causative agent of acute flaccid paralysis (AFP), are positive-sense single-stranded RNA viruses family Picornaviridae. As we approach polio eradication, accurate and timely detection poliovirus in stool from AFP cases becomes vital to success for eradication efforts. Direct PV clinical diagnostic samples using nucleic acid (NA) extraction real-time reverse transcriptase polymerase chain reaction (rRT-PCR) instead current standard method virus isolation culture, eliminates long turn-around time diagnosis need high viral titer amplification laboratories. An essential component direct surveillance is efficient NA. Potential supply issues lack vendor presence certain areas world necessitates validation multiple NA methods. Using retrospective PV-positive (n=104), two kits were compared previously validated Zymo Research Quick-RNA™ Viral Kit. The Roche High Pure Kit, a column-based manual method, MagMaX™ Pathogen RNA/DNA kit used automated Kingfisher Flex system both non-inferior kit, with similar rates pivotal rRT-PCR assays, such as pan-poliovirus (PanPV), serotype 2 (PV2), wild 1 (WPV1). These important assays allow identification differentiation genotypes serotypes fundamental GPLN program. Validation additional provides feasible alternatives piloted assays.