The Alere i Influenza A&B assay is a novel isothermal nucleic acid amplification assay capable of detecting and differentiating influenza A and B viruses in approximately 15 min with minimal hands-on time. This study was conducted in order to assess the performance of the Alere i Influenza A&B assay compared to molecular techniques, considered to be gold standard methods, to evaluate the results. A total of 119 nasopharyngeal swabs collected from inpatients with influenza-like illness were included in the study using both archived and prospectively collected samples from adults and children. Prospectively collected samples were also compared to the Alere BinaxNOW® Influenza A & B Card. The overall sensitivity for detection of influenza A and B viruses compared to those of molecular techniques were 65.96 % and 53.33 % respectively, while the specificity was 98.51 % and 95.96 %. Compared to the Alere BinaxNOW® Influenza A & B Card, the Alere i assay is considerably more sensitive for detection of influenza A and B viruses, although both tests demonstrated excellent specificity for diagnosis of influenza viruses.