Comparative evaluation of RNA isothermal amplification-gold probe lateral flow assay and targeted next-generation sequencing for the detection of Mycoplasma pneumoniae and influenza A and B viruses in children

Objectives: (MP) and Influenza A/B viruses in children with acute respiratory infections (ARIs), and to explore their respective advantages and disadvantages. Methods: Clinical and laboratory data of pediatric patients with ARIs undergoing concurrent RGT and tNGS testing (Jan-Sep 2024) were collected. McNemar's test compared detection rates, and Cohen's kappa coefficient assessed the agreement of the result of between the two methods. Results: For detecting MP and Influenza B virus, tNGS showed a significantly higher positivity rate than RGT. However, there is no difference between tNGS and RGT in the positive detection rate of Influenza A. The agreement between tNGS and RGT was good for MP detection, but only moderate for Influenza A/B virus detection. Conclusions: tNGS offers high-throughput, high-sensitivity screening, while RGT is rapid, user-friendly, cost-effective and superior for identifying active infections and monitoring treatment responses. Optimal method selection is dependent on clinical scenarios and diagnostic priorities.