Clinical Evaluation of the PANDAA Lassa Virus RT-PCR Assay in Sierra Leone

Abstract Molecular diagnosis of Lassa fever is complicated by extensive Lassa virus genetic diversity and limited laboratory capacity in endemic settings. The RealStar© Lassa Virus RT-PCR Kit 2.0 is a commonly used commercial dual-target assay for LASV detection, but its two-target format requires separate amplification reactions, increasing reagent use and workflow complexity. We present the diagnostic evaluation of the single-reaction, adaptive PANDAA Lassa Virus RT-PCR assay with the dual-target RealStar© Lassa Virus RT-PCR Kit 2.0 using 94 archived serum samples from Sierra Leone. 79 of these samples had valid paired results, and we report a positive percent agreement of 78.12% (95% CI 61.25%-88.98%), negative percent agreement of 93.62% (95% CI 82.84%-97.81%), and an overall agreement of 87.34% (95% CI 78.24%-92.98%); Cohen k; was 0.73 (95% CI 0.58-0.89). PANDAA showed substantial agreement and offers a potentially efficient single-well format, but further optimisation and prospective evaluation are needed.