Nanomaterials-based immunochromatographic assays (ICAs) are of great significance in point-of-care testing (POCT), yet it remains challenging to explore low background platforms and high chromogenic intensity probes to improve detection performance. Herein, we reported a low interference and high signal-to-noise ratio fluorescent ICA platform based on ultrabright persistent luminescent nanoparticles (PLNPs) Zn2GeO4: Mn, which could produce intense photoluminescence at 254 nm excitation to reduce background interference from ICA substrates and samples. The prepared immunosensor was successfully applied in T-2 toxin detection with a remarkable limit of detection of 0.025 ng/mL, which was 22-fold more sensitive compared with that of traditional gold nanoparticles. Ultimately, a portable 3D-printed detection device equipped with a smartphone analyzing application was fabricated for quantitative readout in POCT, achieving favorable recoveries in practical sample detection. This work provides a creative attempt for ultrabright PLNP-based low background ICA, and it also guarantees its feasibility in practical POCT.