Abstract: The objective of this study was to determine the replication kinetics of Salmonella enteritidis genome loads (S. enteritidis) in reproductive organs of laying duck after oral challenge. We conducted serovar-specific real-time PCR for S. enteritidis to detect the genomic DNA of S. enteritidis in laying duck at different time points. To validate these results, the Indirect Fluorescent Antibody (IFA) technique was employed too. The results showed that S. enteritidis was consistently detected in all the samples. However, the number of bacteria started decreasing by 3-4 days and by 6 days, the concentration of S. enteritidis DNA was below the detection limits of the PCR assay except the spleen and vagina. In conclusion, this study will help in understanding the pathogenesis of S. enteritidis infection in vivo.