Determining the viable and non-viable load of foodborne pathogens in animal production can be useful in reducing the number of human outbreaks. In this study, we optimized a PMAxx-based qPCR for quantifying viable and non-viable load of from soil collected from free range poultry environment. The optimized nucleic acid extraction method resulted in a significantly higher ( < 0.05) yield and quality of DNA from the pure culture and inoculated soil samples. The optimized primer for the amplification of the gene fragment showed high target specificity and a minimum detection limit of 10 viable from soil samples. To test the optimized PMAxx-based qPCR assay, soil obtained from a free range farm was inoculated with Enteritidis or Typhimurium, incubated at 5, 25, and 37°C over 6 weeks. The survivability of Typhimurium was significantly higher than Enteritidis. Both the serovars showed moisture level dependent survivability, which was significantly higher at 5°C compared with 25°C and 37°C. The PMAxx-based qPCR was more sensitive in quantifying the viable load compared to the culture method used in the study. Data obtained in the current study demonstrated that the optimized PMAxx-based qPCR is a suitable assay for quantification of a viable and non-viable load of from poultry environment. The developed assay has applicability in poultry diagnostics for determining the load of important serovars containing .