Surface plasmon resonance (SPR) as a rapid tool for serotyping of Salmonella.

An SPR-based sandwich immunoassay for serotyping of Salmonella is demonstrated. The Salmonella are captured on an SPR chip using polyclonal capture antibody. SPR sensorgrams are obtained for the immunoreactions of the somatic (O) and flagellar (H) surface antigens, of the captured bacteria, to their respective antibodies. The sensorgram data are compiled to determine the antigenic formula in accordance with the Kauffmann-White scheme. Salmonella Enteritidis was completely serotyped using this SPR-based method. In addition, Salmonella belonging to serogroups B, C and D were successfully assigned to their respective serogroups. Before serotyping the bacteria are grown to a concentration of 1x10(10) mL(-1). This SPR-based serotyping provides quantitative data, and thus, eliminates the possibility of false detections as encountered in the conventional slide agglutination test (SAT). This method was also proved to work with rough strains.

A rapid serological assay for prediction of Salmonella infection status in slaughter pigs using surface plasmon resonance.

We present a rapid surface plasmon resonance-based serological assay for the detection of Salmonella Typhimurium infection in pigs using the Plasmonic((R)) SPR device. Lipopolysaccharide (LPS, 10 microg mL(-1)) from Salmonella Typhimurium was immobilised by self-assembly on a hydrophobic SPR chip. Using this LPS-coated chip, it was possible to bind and detect the anti-Salmonella Typhimurium antibodies in serum of pigs infected with the bacteria. The developed SPR assay is able to differentiate between sera obtained from pigs having low, medium, and high levels of Salmonella infection. A commercial ELISA kit was used to classify the sera for levels of Salmonella infection on the basis of optical density (OD%). A strong positive correlation was observed between the SPR-based assay and the ELISA (n=38, r=0.90, p<0.01). The sensitivity and specificity of the assay are 0.93 and 0.87, respectively. The SPR-based assay is label-free and does not require any sample preparation or dilution steps. The total analysis time is 45 min for each serum sample. The assay was found to be specific for Salmonella Typhimurium and shows no cross-reactivity to Salmonella Choleraesuis or Escherichia coli antibodies. As no sample preparation is required the developed assay has the potential to be used as a reliable tool for Salmonella monitoring programmes in pork production.

Rapid method for detection of Salmonella in milk by surface plasmon resonance (SPR).

The Plasmonic surface plasmon resonance (SPR) device was used to develop a rapid, simple and specific immunoassay for detection of Salmonella in milk. Rapid detection of Salmonella contamination is a major challenge for the food industry. Salmonella contamination is well known in all foods including pasteurised milk. The SPR assay was developed as a sandwich model using a polyclonal antibody against Salmonella as capture and detection antibody. Milk spiked with Salmonella typhimurium cells, killed by thimerosal (1%, w/w) treatment was used. Using the Plasmonic SPR assay it was possible to detect S. typhimurium down to a concentration of 1.25 x 10(5) cells ml(-1) in both milk and buffer system. The results obtained are comparable with existing, approved rapid Salmonella detection techniques. No negative effects on the sensitivity of the assay are encountered due to the milk matrix. Hence, no sample preparation or clean-up steps are required. The sample volume requirement for the assay is only 10 microl. Using the assay S. typhimurium was detected in milk within 1h, whereas the cultural techniques require 3-4 days for presumptive positive isolates and further time for confirmation. The rapid tests require at least 24h for the results. The Plasmonic SPR device operates on the Kretschmann configuration and is a cuvette-based system with the advantage of having eight channels on one single SPR chip.