Rapid Isothermal detection and Quantification of Total Salmonella in Poultry Using the HyperKit Point-of-Use diagnostic test

Food producers face mounting pressure to ensure both safety and efficiency as food systems expand in scale and complexity. However, existing diagnostic tools often force a trade-off between speed, accuracy, cost, and usability, leaving the industry with limited options for real-time, on-site pathogen detection. This study evaluated the performance of the HyperKit Total Salmonella (HK), a novel point-of-use rapid diagnostic tool for detecting and semi-quantifying Salmonella in artificially-spiked and commercial chicken primal samples. HK is composed of one-step sample processing tool and a room temperature fluorescent master mix. Targeted DNA was amplified at 65 ^o C for 60 min and fluorescence measure over time at 495 nm. HK successfully detected Salmonella in all spiked samples (1.0 to 7.5-log CFU/mL; n = 57) under 60 minutes. HK demonstrated high semi-quantitative accuracy (r² = 0.93; P < 0.001), particularly at concentrations ≥ 1.5 log CFU/mL (± 0.18-log precision), as well as strong repeatability (0.16-log; 95% CI: 0.11–0.21) and reproducibility across three operators and samples of multiple origins (Georgia, Illinois, Nebraska). Data obtained with HK were in close agreement with the reference microbiology method, but up to 300 times faster. Robustness studies confirmed reliable performance under varying sample preparation conditions. Importantly, the kit showed complete inclusivity for all tested Salmonella serotypes (n = 46) and strong exclusivity against non-target organisms (n = 37). In conclusion, this study demonstrated that HK is a rapid and accurate detection tool with a simple field workflow to support point-of-use applications in food processing environments, and thus, enhancing food safety monitoring and response in decentralized settings.