Rapid Detection of Salmonella Enterica Serovar Enteritidis from Eggs and Chicken Meat by Real-Time Recombinase Polymerase Amplification in Comparison with the Two-Step Real-Time PCR

Salmonella is the most common cause of foodborne outbreaks throughout the world and Salmonella enterica serovar Enteritidis is one of the major causes of salmonellosis. We describe a rapid, sensitive and inexpensive method for the detection of S. Enteritidis from eggs and chicken meat using the real‐time recombinase polymerase amplification (Real‐time RPA) and compared this assay with the two‐step real‐time polymerase chain reaction (PCR) (Real‐time PCR). The detection sensitivity of the RPA and PCR assays was identical in pure culture, however, the reaction time was as short as 10 min compared with 40 min for the PCR assay. In food application, the detection sensitivity of the RPA assay was 10 cfu/g from eggs and 100 cfu/g from chicken samples without additional DNA purification steps before the amplification – therefore, the sensitivity of the RPA assay was 100 times greater from eggs and 10 times greater from chicken than the PCR assay. Practical Applications Numerous epidemiological investigations attributed the source of sporadic Salmonella outbreaks as poultry and poultry‐by‐products including eggs. This is the first study in which a real‐time RPA was developed to detect the Salmonella Enteritidis specifically. The whole procedure is fairly simple and does not require specific equipment, making it potentially applicable in the monitoring and detection of S. Enteritidis in eggs and poultry products.