Rapid TRC for Influenza Detection

For TRCsatFLU, we soaked a new swab in a gargle sample for 5 s, and the swab containing the gargle solution was mixed with 1 mL extraction buffer containing surfactant, and it was infected into a single-use cartridge that contains all the elements required for rapid TRC. Nasopharyngeal swabs were mixed with 1 mL extraction buffer containing surfactant. Of the extraction buffer, 140 μL was aliquoted for RT-PCR, and the remaining 860 μL was injected into the TRCsatFLU cartridge. Next, the cartridge was set in TRCsat®, and nucleic acid amplification, detection, and determination of results were automatically performed in the instrument. The procedure of rapid TRC in TRCsat® is as follows: the samples were incubated at 52 °C for 1 min and then mixed 30 μL of the sample with a dry reagent containing enzymes, substrates, primers, and INAF probes and incubated at 46 °C to monitor fluorescence; it was automatically determined as positive when the fluorescence intensity ratio of the reaction solution exceeded 1.2. TRCsatFLU was designed to detect Influenza A(H1N1), A(H1N1)pdm09, A (H3N2), and B (Victoria and Yamagata lineages), but TRCsat® only showed the results with influenza A or B positive or influenza negative.