A Highly Sensitive ELISA and Immunochromatographic Strip for the Detection of Salmonella typhimurium in Milk Samples

Abstract

Murine monoclonal antibodies to target Salmonella typhimurium flagellin and lipopolysaccharide (LPS) were prepared and characterized. For the immunological detection of S. typhimurium, different pairs of monoclonal antibodies (MAbs) were tested in a sandwich enzyme linked immunosorbent assay (ELISA) format. After comparison, a sandwich ELISA and immunochromatographic strip based on LPS MAbs was established to detect S. typhimurium. The determination limits of the immunochromatographic strip in phosphate-buffered saline (PBS) containing 0.1% Tween 20 (PBST) and pure milk sample were found to be 1.25 × 10⁵ colony-forming units (cfu)/mL and 1.25 × 10⁶ cfu/mL S. typhimurium, respectively. Results can be obtained with the naked eye in 10 min. Cross-reactivity was observed with Salmonella paratyphi B, but not S. paratyphi A or Salmonella enteritidis. The LPS MAbs based immunochromatographic strip is rapid and convenient to detect S. typhimurium in milk samples.