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Photostable methylene blue‐loaded silica particles used as label for immunosorbent assay of Salmonella Typhimurium
Author(s) -
Cheeveewattanagul Nopchulee,
Tien Tran Thanh,
Rijiravanich Patsamon,
Surareungchai Werasak,
Somasundrum Mithran
Publication year - 2019
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1796
Subject(s) - salmonella , methylene blue , chromatography , chemistry , detection limit , polyclonal antibodies , analyte , immunoassay , absorbance , bioconjugation , enzyme , monoclonal antibody , enterobacteriaceae , colorimetry , microbiology and biotechnology , bacteria , biochemistry , antibody , escherichia coli , biology , genetics , photocatalysis , gene , immunology , catalysis
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 598 nm. The platform had an assay time of 20 min, could detect heat‐killed Salmonella with a limit of detection of 48 CFU mL −1 , and gave good recoveries in milk. The labels could be stored at 4 °C for 70 days without any deterioration.