Food allergy has become an important problem affecting people's health. As one of the most common allergenic food, crustaceans pose a dietary risk to allergic sensitizers. A rapid and sensitive detection method is needed for allergen monitoring. We developed a quantum-dot-based fluorescent lateral flow Immunoassay (LFIA) for detection of major crustacean allergen tropomyosin (TM). Polyclonal antibodies were labeled by quantum dots through covalent bonding and characteristics of combined probe were analyzed. Relevant conditions of strip preparation were optimized, with results of 0.8 mg/mL test line coating concentration and 1% Tween-20 10 mM PBS as flowing buffer. The detection limits of developed assay were 0.5 μg/mL visually and 0.05 μg/mL by instrument without complex operation. The LFIA showed good specificity and reproducibility, and the results for real food samples were consistent and comparable to validated ELISA kit. The novel developed detection strip would significantly improve applications for the on-site detection of TM in foods.