Seed-mediated synthesis of gold nanorods (AuNRs) has been widely used for diverse applications in the past decade. In this work, this synthetic process is demonstrated for multicolor biosensing for the first time. Our investigation reveals that ascorbic acid acts as a key factor to mediate the growth of AuNRs. This phenomenon is incorporated into the alkaline phosphatase (ALP)-enzyme-linked immunosorbent assay (ELISA) system based on the fact that ALP can catalyze the conversion of ascorbic acid-phosphate into ascorbic acid with high efficiency. This allows us to develop a multicolor ELISA approach for sensitive detection of disease biomarkers with the naked eye. We show the proof-of-concept multicolor ELISA for the detection of prostate-specific antigen (PSA) in human serum. The results show that different colors are presented in response to different concentrations of PSA, and a detection limit of 3 × 10(-15) g mL(-1) in human serum was achieved. The proposed multicolor ELISA could be a good supplement to conventional ELISA for POC diagnostics.

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