Development of a nanostructured immunosensor for early and in situ detection of Xanthomonas arboricola in agricultural food production.

We report a microfluidic electrochemical immunosensor for Xanthomonas arboricola (XA) determination, based on the covalently immobilization of monoclonal anti-XA antibody (anti-XA) on a previously amino functionalized SBA-15 in situ synthesized in the central channel of a glass-poly(dimethylsiloxane) microfluidic immunosensor. The synthetized amino-SBA-15 was characterized by N2 adsorption-desorption isotherm, scanning electron microscopy and infrared spectroscopy. XA was detected by a direct sandwich immunoassay through an alkaline phosphatase (AP) enzyme-labeled anti-XA conjugate. Later, the substrate p-aminophenyl phosphate was converted to p-aminophenol by AP. The enzymatic product was detected at +100mV on a sputtered gold electrode. The measured current was directly proportional to the level of XA in walnut trees samples. The linear range was from 5 × 102 to 1 × 104CFUmL-1. The detection limit was 1.5 × 102CFUmL-1, and the within- and between-assay coefficients of variation were below 5%. Microfluidic immunosensor is a very promising tool for the early and in situ diagnosis of XA in walnuts avoiding serious economic losses.