Molecular study of amoebopore-c gene of entamoebahistolytica isolated from food handlers in Erbil city

Background and objective: Entamoebahistolytica is a protozoan parasite with high prevalence rates in tropical and subtropical regions of underdeveloped countries. The outcomes of Entamoebahistolytica infection are mostly asymptomatic; only a fraction of those infected develop dysentery and, in rare cases, liver abscesses. Its pathogenicity is due to a huge number of virulence factors. This study investigated the risk of Entamoebahistolytica among food handlers, screening, Sequencing, and phylogenetic tree of Ameobopore-c. Methods: Total of 563 stool samples were collected from food handlers who visit the central laboratory in Erbil City every year. Wet mount microscopic examination was performed to diagnose the Entamoeba species, molecular analysis was done for positive samples. FavorPrep Stool for DNA Isolation Mini Kit (Favorgen, Taiwan) was utilized. PCR analysis was done targeting the Amoebopore-c gene using one set of primers to amplify the 705 bp fragment. Gel electrophoresis was performed to visualize the amplified DNA under Ultraviolet light. Fifteen random positive samples of the PCR product were sent to Macrogen in South Korea to determine the DNA sequence of the Amoebopore-C gene present in the Entamoebahistolytica. Results: The study found no statistically significant relationships between infection rates and demographic or socioeconomic characteristics. In fact, that 21 of the 50 microscopic positive cases were also positive by PCR.15 sequences of the amoebapore-C gene revealed that a modification in a single nucleotide leads to a modification in a single amino acid. Alignment of amoebapore C, according to NCBI-BLAST Homology Sequence, showed the following identity percentages to samples from different countries: India (99.4%), the USA (99.18% and 99.7%), Iraq (99.38%), and Hamburg (99.84%) Conclusion: PCR is a highly sensitive method for detecting Entamoebahistolytica. The Entamoebahistolytica isolates showed high homology in amoebapore-C gene sequences compared to global strains.