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^TM-based qPCR for quantifying viable and non-viable load of Salmonella from soil collected from free range poultry environment. The optimized nucleic acid extraction method resulted in a significantly higher (P < 0.05) yield and quality of DNA from the pure culture and Salmonella inoculated soil samples. The optimized primer for the amplification of the invA gene fragment showed high target specificity and a minimum detection limit of 10² viable Salmonella from soil samples. To test the optimized PMAxx^TM-based qPCR assay, soil obtained from a free range farm was inoculated with Salmonella Enteritidis or Salmonella Typhimurium, incubated at 5, 25, and 37°C over 6 weeks. The survivability of Salmonella Typhimurium was significantly higher than Salmonella 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^TM-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^TM-based qPCR is a suitable assay for quantification of a viable and non-viable load of Salmonella from poultry environment. The developed assay has applicability in poultry diagnostics for determining the load of important Salmonella serovars containing invA.

确定动物生产中食源性病原体的可行和不可行载量可用于减少人类暴发的数量。在这项研究中，我们优化了基于PMAxx ^TM的qPCR，用于定量检测沙门氏菌从自由放养的家禽环境中收集的土壤中。优化的核酸提取方法可显着提高P <0.05）来自纯培养物的DNA的产量和质量 沙门氏菌接种土壤样品。优化的引物用于扩增invA基因片段显示出高靶标特异性，最低检测限为10 ^2有活力沙门氏菌从土壤样品中提取。为了测试基于PMAxx ^TM的优化qPCR分析方法，对从自由放养场获得的土壤进行了接种沙门氏菌 肠炎或 沙门氏菌鼠伤寒，在5、25和37°C下孵育6周。的生存能力沙门氏菌 鼠伤寒明显高于 沙门氏菌肠炎。两种血清素均显示出湿度水平依赖性的生存能力，与25°C和37°C相比，在5°C时显着更高。与研究中使用的培养方法相比，基于PMAxx ^TM的qPCR在定量可行负荷方面更为敏感。当前研究中获得的数据表明，基于PMAxx ^TM的优化qPCR是用于定量检测是否存在活菌的合适方法。沙门氏菌来自家禽环境。所开发的测定方法可用于家禽诊断中确定重要的负荷沙门氏菌 含有 invA。