1：Experimental Design and Method Selection:

The study designed a portable TFT biosensor with a dual-gate structure for enhanced sensitivity, packaged to protect against environmental factors. It utilized electrochemical detection based on antigen-antibody reactions.

2：Sample Selection and Data Sources:

Samples included oral and cloacal swabs and feces from ducks and mallards infected with HPAIV H5N6, prepared in a BSL-3 laboratory. Virus concentrations were determined using RT-PCR for comparison.

3：List of Experimental Equipment and Materials:

Materials included glutaraldehyde, ethanolamine, N-hydroxysuccinimide, bovine serum albumin, 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide hydrochloride, lysis buffer (from Bionote rapid test kit RB2501MH), HPAIV antibodies, and various buffers. Equipment included a packaged biosensor with TFT device, internal and external PCBs, gold wires, epoxy for sealing, RF magnetron sputter for tin dioxide deposition, O2 plasma system, and a current-voltage sourcemeter for measurements.

4：Experimental Procedures and Operational Workflow:

The biosensor was fabricated by wiring TFT devices on PCBs and sealing with epoxy. Antibodies were fixed to the sensing gate surface using chemical functionalization. Samples were reacted with antibodies for 20 minutes, washed, and measured for voltage shifts. Measurements were conducted in a BSL-3 environment with controlled parameters (drain voltage of 6 V, bottom gate sweep from -10 to +20 V).

5：Data Analysis Methods:

Voltage shifts were calculated as the difference between reference and sample measurements. Data were compared with RT-PCR results to establish correlation, and statistical analysis included linear fitting and error calculation.