Viruses such as influenza virus (Flu), respiratory syncytial virus (RSV), and SARS-CoV-2 are the major cause of acute respiratory infections (ARIs) in children, resulting in significant morbidity and mortality worldwide [1], [2], [3]. CDC estimated 12 million flu-related illnesses, 45,000 hospitalizations and 550 flu related deaths in children in the US during 2023–24 [4]. RSV led to 2.1 million outpatient visits and 58,000–80,000 hospitalizations annually among children less than 5 years of age in the US [5]. COVID-19 accounted for approximately 234,000 hospitalizations in children < 18 years of age in the US between fall 2020 to spring 2024 [6]. Pediatric ARI is also associated with significant healthcare burden accounting for approximately 300 outpatient visits/1000 children and adolescents each year in the US [7]. Usually self-limiting, viral ARIs often resolve with over-the-counter remedies. However, an estimated 21 % of outpatient antibiotics prescribed for pediatrics in the US are inappropriate and are for a viral ARI [8]. This overuse results in an increased risk of medication-induced adverse effects and associated annual national costs [9]. Due to the heavy burden of viral ARI, an early and accurate detection and differentiation of these vital pathogens by sample-to-answer, molecular point-of-care (POC) tests is needed for effective triage, discontinuation of antibiotics, and timely implementation of infection control measures.

The BioFire SPOTFIRE Respiratory/Sore Throat Panel (SPOTFIRE), (BioFire Diagnostics, bioMérieux) detects and identifies 11 viruses and 4 bacteria causing respiratory illness (Supplementary Table 1) or pharyngitis from nasopharyngeal swabs (NPS) or throat swabs, respectively, within 16 min turnaround time (TAT) [10]. Xpert® Xpress SARS-CoV-2/Flu/RSV PCR assay (Xpress) (Cepheid, Sunnyvale, CA) detects SARS-CoV-2, influenza A and B viruses and RSV A/ B in NPS or anterior nasal swabs within a runtime of about 40 min [11]. Cobas® SARS-CoV-2 & Influenza A/B assay (Liat) (Roche Diagnostics, USA) detects SARS-CoV-2, Flu A and Flu B in both NPS and nasal swabs in 20 min [12], [13].

Previous studies have compared the detection of these viruses with Xpert® Xpress and Liat® assays [12], [13], [14], [15], [16], [17]; however, clinical evaluation of SPOTFIRE is limited. A single study from Hong Kong demonstrated comparable performance of SPOTFIRE to BioFire FilmArray Respiratory 2.1 plus Panel [10] but there is no data to compare the performance of this assay with other common multiplex POC assays. We evaluated SPOTFIRE for detection of respiratory viruses from pediatric NPS and compare its performance to Xpress assay for SARS-CoV-2/Flu/RSV and Liat assay for SARS-CoV-2 & Influenza A/B.