Exosomes are Extracellular Vesicles (EV) that own unique structural features and functions and have gradually become the hot research spot in recent years. The tumor-derived exosomes contain various types of useful biological information, and medical identification of exosomes relied on the specific characterization of membrane surface proteins. In this study, in order to rapidly identify non-small cell lung cancer (NSCLC)-derived exosomes, based on an aptamer against CD63 protein on exosome membrane, a low cost lateral flow aptamer assay (LFAA) test strip using nanogold particles as visualization probes was successfully developed for facile identification of A549 exosomes isolated from human lung carcinoma cells diluted from 6.4 × 10 particles/mL herein.