Improved bioluminescent enzyme immunoassay for the rapid detection of Salmonella in chicken meat samples

Abstract

S . F U K U D A , H . T A T S U M I , S . I G I M I A N D S . Y A M A M O T O . 2005.Aims: To evaluate an improved bioluminescent enzyme immunoassay (BEIA) using biotinylated firefly luciferase for the rapid detection of Salmonella in naturally contaminated chicken meat samples. Methods and Results: Capture agents and lipopolysaccharide (LPS) extraction reagents for Salmonella were investigated to improve the sensitivity of the BEIA. Also, the use of Oxoid SPRINT (Simple Pre-enrichment and Rapid Isolation New Technology) as a pre-enrichment and selective medium for 26-h BEIA detection of Salmonella in chicken meat samples was examined. The use of polymyxin B as a capture agent on solid support and 3-[(3-Cholamidopropyl) dimethylammonio] propanesulfonic acid (CHAPS) for extraction of the LPS facilitated sensitive detection of Salmonella. Of 120 chicken meat samples, 25 samples were positive using the improved BEIA with the SPRINT and 25 samples were positive using the SPRINT followed by the standard isolation methods. Conclusions: The improved BEIA, in which polymxin B was used as a capture agent and CHAPS was used for extraction of the antigen, had a sensitivity of 96AE0% and a specificity of 98AE9% for the detection of Salmonella in chicken meat. Significance and Impact of the Study: The improved BEIA combined with the SPRINT medium for the detection of Salmonella in chicken meat samples produced comparable results to the culture methods in 26 h.

Firefly Luciferase‐based Fusion Proteins and their Applications in Bioanalysis

“…Complexes of biotinylated luciferase with streptavidin are also used for detection of pathogenic microorganisms and their surface antigens (protein A, protein G , as well as lipopolysaccharides ), and for differentiation of strains differing in resistance to antibiotics . Despite the high sensitivity of detection of the luciferase probe, the minimum detecting concentration of cells was found to be rather high in the range of ~1 × 10 4 ‐5 × 10 5 CFU mL −1 .…”

“…For example, Fukuda and co‐authors detected the presence of ~50 CFU mL −1 of Staphylococcus aureus cells after 7 h of cultivation . A method for 24‐h bioluminescent immunoassay of Salmonella in chicken meat was suggested in , and the cultivation medium composition was optimized in together with the capturing agent immobilized on the surface of the plate.…”

Immunoassay based on carbon nanotubes-enhanced ELISA for Salmonella enterica serovar Typhimurium

“…Nevertheless, the detection sensitivity was considered close to that of a conventional ELISA (10 −5 -10 −6 CFU/ml). In addition, the firefly luciferase was also used to replace enzyme in a technique called bioluminescent enzyme immunoassay (BEIA) (Fukuda et al, 2005). In this case, in order to lower the detection limit, a specific type of culture media must be used.…”

Immunological Methods for the Detection of Campylobacter spp. - Current Applications and Potential Use in Biosensors

“…The limitation of this method is that treatment of bacteria with zwitterionic detergents is necessary (Blais 2005(Blais , 2006Brooks et al, 1998). Fukuda et al (2005 ) demonstrated that PMB capture was superior to antibody capture for detection of Salmonella in chicken. Our laboratory demonstrated that PMB-capture enhances ELISA detection of C. jejuni as indicated by a 10-fold improvement in the limits of detection with both the USB10 and USB14 antibodies (unpublished results).…”