Novel dual-gene target and probe-based real-time PCR for the detection and differentiation of <i>Salmonella</i> Dublin

serovars. Unlike many current tools, this assay is validated for direct use with challenging samples, including feces, nasal and vaginal swabs, and environmental boot swabs, where inhibitors and low pathogen load commonly interfere with detection. By enhancing sensitivity and reliability across diverse matrices, this method supports farm-level surveillance, rapid outbreak response, and risk-based control strategies. Broader implementation can improve herd health, limit economic losses, and reduce the risk of human exposure through food or environmental sources.