Rapid and sensitive detection of Salmonella spp. in raw minced meat samples using droplet digital PCR

Abstract

With its ability to easily infect each step of food production chain, Salmonella spp. stands as one of the most important foodborne pathogens which impose threats against economy and human health by creating high morbidity and mortality, worldwide. Although sensitive screening and detection methodologies based on common conventional techniques have been developed for Salmonella spp., their long turnaround time and strict legislative demands directs the researchers to develop more sensitive and rapid methods. Digital PCR has many potential advantages over its closest competitor, Real-Time PCR. It gives more accurate, sensitive, and precise results, especially for samples that contain low pathogen concentrations and shows more resistance against PCR inhibitors. In this study, we developed a droplet digital PCR screening methodology with a short pre-enrichment step. For this purpose, 25 g raw minced meat was subjected to Salmonella contamination, and following a very short 3 h enrichment step, bacterial isolation from 5 mL of sample was achieved. For the specific detection of the Salmonella spp., bipA gene region was selected as a better candidate. Subsequently, droplet digital PCR setup was performed and detection sensitivity of the test was determined as 1.39 cfu/mL. Furthermore, parallel experiments were also conducted using Real-Time PCR, for comparative and validative purposes. Conclusively, data gathered in this study have shown that droplet digital PCR with our proposed setup can sensitively detect Salmonella spp. in raw minced meat samples while generating the same-day results.