An HRP-labeled lateral flow immunoassay for rapid simultaneous detection and differentiation of influenza A and B viruses

Rapid and sensitive diagnosis of influenza is urgently needed to address the limitations of low sensitivity associated with current rapid tests available for clinics and on-site monitoring. A novel horseradish peroxidase (HRP)-labeled lateral flow immunoassay strip (HRP-LFIA) for rapid simultaneous detection and differentiation of influenza A (INF A) and influenza B (INF B) viruses were developed. This immunoassay was based on the signal amplification by the HRP-catalyzed oxidation of 3, 3', 5, 5'-tetramethylbenzidine forming a colored insoluble product, which was proportional to the analyte concentration. Compared with conventional gold-colloidal based strips, an analytical sensitivity enhancement of more than one order of magnitude for thirteen INF virus isolates was observed. A total of 1487 swabs obtained from persons with influenza-like illnesses were tested for the presence of INF A and B viruses using real-time reverse transcription polymerase chain reaction (rRT-PCR) as the reference criterion. The overall sensitivities of HRP-LFIA were 77.5% (100/129) and 71.2% (116/163) for INF A and INF B, respectively. The overall specificities were 99.8% (1144/1146) and 99.8% (918/920), respectively. The nasopharyngeal sampling method yielded higher sensitivity rates of 90.2% (55/61) and 82.6% (71/86). In conclusion, this user-friendly assay could be a promising rapid detection method for rapid screening of INF A and INF B viruses.