Photostable methylene blue-loaded silica particles used as label for immunosorbent assay of Salmonella Typhimurium.

Abstract

Salmonella Typhimurium is a major cause of food poisoning. To solve the limitations of the routine enzyme linked immunosorbent assay such as laborious assay procedure, lack of long-term enzyme stability, and insufficient sensitivity, we provided a non-enzymatic colorimetric immunosorbent assay platform to overcome these problems. The highly photostable redox dye particles was constructed by silica particles (diameter = 598±14.4 nm) loaded with methylene blue (Si-MB) and applied to be a label for immunoassay of S. Typhimurium. The sandwich assay format involved incubation of an analyte in a microplate wells modified with monoclonal anti-Salmonella, followed by exposure to a polyclonal anti-Salmonella/Si-MB bioconjugate and then measurement of absorbance at 598nm. The platform had an assay time of 20min, could detect heat-killed Salmonella with a limit of detection of 48CFU mL-1 , and gave good recoveries in milk. The labels could be stored at 4°C for 70 days without any deterioration.