Multicenter evaluation of Coyote FlashDetect LyocartE multiplex assays for six respiratory pathogens: a comparative analysis with Sansure and Cepheid assays

Abstract

ABSTRACT Respiratory diseases caused by common respiratory pathogens, such as influenza A/B (Flu A/B), severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), respiratory syncytial virus (RSV), adenovirus (ADV), and Mycoplasma pneumoniae (MP), share many of the same symptoms and may progress to severe complications in some cases. Multiplex molecular point-of-care testing (mPOCT) enables automated and rapid differential diagnosis for timely intervention. This study independently evaluated the clinical performance of Coyote FlashDetect LyocartE SARS-CoV-2/FluA/FluB and FlashDetect LyocartE RSV/MP/ADV multiplex mPOCT assays using the Sansure as the primary comparator, followed by a subset-based, target-specific paired comparison with Cepheid mPOCT assay for selected respiratory pathogens (FluA, FluB, and RSV). Specifically, 642 and 427 throat swab specimens were used to evaluate the Coyote SARS-CoV-2/FluA/FluB and Coyote RSV/MP/ADV multiplex mPOCT assays against Sansure assays, respectively. Coyote assays demonstrated high concordance with Sansure assays, with overall percent agreement (OPA) values ranging from 93.8% to 100.0% for six pathogens covered by two Coyote assays. Most discrepancies with Sansure assays occurred in samples with cycle threshold (Ct) values ≥35, especially among FluA samples. Moreover, 414, 414, and 215 specimens were allocated for FluA, FluB, and RSV, respectively, to conduct target-specific paired comparison between Coyote and Cepheid. Each Coyote assay also demonstrated high concordance with the Cepheid assays, with PPA values of 96.0%, 100.0%, and 98.3% for FluA, FluB, and RSV, respectively. These results support the use of Coyote mPOCT assays in specific settings. Coyote assays demonstrated a high degree of concordance for shared targets with comparator methods, supporting their use as a rapid and user-friendly diagnostic approach for multiplex respiratory pathogen detection. IMPORTANCE This study highlights the value of multiplex molecular point-of-care tests. The POCT assays demonstrate high concordance with the laboratory-based lab tests in this retrospective specimen cohort, offering a viable, rapid, and user-friendly alternative for multiplex respiratory pathogen detection and hold the promise to be used in primary healthcare settings and public health surveillance.