Preliminary Development of a Competitive Fluorescent Quantum Dot-Based Immunochromatographic Test Strip for Sensitive Saxitoxin Detection

Abstract

Rapid and portable detection of saxitoxin (STX) and its many congeners is highly desirable to prevent paralytic shellfish poisoning due to red tide or harmful algal blooms. In this work, we describe successful preliminary efforts to develop a very sensitive general STX family test strip employing highly fluorescent red quantum dots (Qdot 655) to detect as little as 0.5 to 1 part per billion (ppb or ng/ml) of STX with a dynamic range extending to 20,000 ppb after the prototype dipstick assay was optimized. A competitive format was necessitated by the small molecule nature of STXs having only one epitope, but the decrease in Qdot fluorescence was clearly visible to the naked eye as a function of increasing STX concentration in aqueous buffer. The competitive displacement assay format required conjugation of a primary amine in STX to carboxyl-Qdot 655 via a covalent carbodiimide coupling reaction which was validated by an electrophoretic mobility band shift assay.