Salmonella is one of the most common causes of foodborne outbreaks and infection worldwide. The gold-standard detection method of Salmonella is cultivation. There is a need to investigate rapid and accurate processes with time-consuming cultivation. The study evaluated diferent approaches to detect Salmonella in poultry feces samples. Poultry farm feces samples from 21 cities in Iran were collected from January 2016 to December 2019. Microbiological cultures, serological assays, and multiplex PCR (m-PCR) were used to detect and characterize Salmonella spp. isolates. Serological assays and m-PCR were used to determine the serogroups A, B, C1, C2, D1, E, H, and FliC. The m-PCR was used to detect seven Salmonella serovars, and a Chi-square test was performed to compare the discriminatory power of the methods. Of 2300 poultry feces samples, 173 (7.5%) and 166 (7.2%) samples were detected as Salmonella spp. by cultivation and m-PCR, respectively. The sensitivity of the molecular method was equal to cultivation at 0.96 (CI=95%). Assessment of H antigenic subgroups showed the same for both m-PCR and serological tests. Therefore, the matching rate of the two methods for detecting all H antigenic subgroups was 100%. Thus, the relationship between the results obtained from both methods was signifcant in the contingency table test (P<0.01). The PCR-based approach confrmed the detection of Salmonella in a shorter period (24–36 h) compared to the conventional microbiological approach (3–8 days)
