The rapid and accurate detection and enumeration of low levels of Salmonella Typhimurium in food processing facilities are critical components of an effective hazard analysis critical control point program. The objective of this study was to develop a rapid (8 h) most probable number (MPN)-enzyme-linked immunosorbent assay (ELISA) for the detection and enumeration of Salmonella Typhimurium in wastewater. The specific objectives were to (i) characterize poly- and monoclonal Salmonella Typhimurium-specific antibodies in order to select the most specific and sensitive antibody for Salmonella Typhimurium detection, and (ii) validate the MPN assay through a correlation between the 8-h MPN-ELISA and the traditional 48-h Salmonella Typhimurium MPN method in poultry scald water. Poultry scald water samples were spiked with 10 and 50 CFU/ml of Salmonella Typhimurium. The traditional MPN method used a 48-h enrichment period followed by an analysis, while the MPN-ELISA used a 5-h enrichment period followed by a 3-h ELISA analysis. No differences (P < 0.05) were found between the traditional MPN and the MPN-ELISA, indicating the promise of the MPN-ELISA for the rapid detection and enumeration of Salmonella Typhimurium within an 8-h shift. This abbreviated assay will permit increased product sampling and more rapid movement of food between production and processing, resulting in reduced spoilage and quality losses.