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This investigation was done for isolation and molecular identification of non-typhoidal salmonella from humans. From 2021 till end of 2022, 100 stool samples were collected from humans with diarrhea different clinics of Baghdad province. After 24 hours of incubation at 37 °C, the samples were grown on several selective medium to identify salmonella colonies. Multiple biochemical analyses were performed on the growth isolates, and the results were verified using the Api20-E system. Colonies having biochemical properties consistent with Salmonella spp. were examined using the API20-E method, and their identities were confirmed using PCR targeting 16S rRNA. Six (6%) human Salmonella entericaserovars isolates were positively subspecies-identified by routine bacteriological method. In this work, the 16S rRNA gene was used to screen for Salmonella spp. in isolates from diarrhoea samples. A total of 6 S. enterica isolates were taken from people, and 6 (6%) of those isolates showed positive amplifications in the first round of PCR, which included running the 16S rRNA 1500bp gene.