The current study was for determine Entamoeba dispar & Entamoeba moshkovskii in symptomatic individuates by molecular identification ( nested  multiplex PCR ) in order obtained  for an  accurate  diagnosis of  these Entamoeba species. Total stool samples (120) that collected from symptomatic patients with diarrhea. These samples were collected from patients according to age, residency during the period from ( the first of November 2021 to the end of January 2022). All samples undergo for a direct microscopic examination, The results  revealed that 100(73.18%)were positive samples with Entamoeba spp. The results of microscopic examination recorded high infection rate in age group in (4-10)years83.33%. In contrast, recorded high prevalence in rural areas 76.31% than urban areas 73.68% . Showed the method nested multiplex polymerase chain reaction by using   18s  rRNA small subunit gene, the first round  of PCR showed the infection rate by Entamoeba spp 87(65.41%) out of 120 stool samples .The second round of nested multiplex PCR for these positive samples by using  two specific primers revealed mix infection of Entamoeba dispar & Entamoeba moshkovskii with Entamoeba histolytica   5(6.25%), 2(2.5%) respectively, with significant difference at P<0.05.

The current study was for determine Entamoeba histolytica in diarrheal patients by molecular investigation (nested multiplex PCR) in order obtained for an accurate diagnosis of Entamoeba histolytica that causes amoebiasis. Total stool samples (133) that collected from symptomatic patients with diarrhea according to age, gender. Then all samples undergo for a direct microscopic examination, The result revealed that 100(73.18%) were positive samples with Entamoeba spp. The results of microscopic examination revealed a higher infection rate of Entamoeba spp were in the female 83.87% compared the male 67.6% with significant difference at P<0.05 ,and recorded high infection rate in age group in (4-10)years and age group (more than 50)years  83.33%,86.66% respectively, and lower infection rate in age group (>1)year 55.55% .Showed the method nested multiplex polymerase chain reaction by using 18s rRNA small subunit gene, nested multiplex PCR for these positive samples by using  specific primer for E. histolytica revealed a higher infection rate was E. histolytica 73(91.25%) (monoinfection). Entamoeba histolytica   high infection rate recorded in the female 97.43% compared the male 82.35%. In addition, E. histolytica recorded the more percentage infection was in age group (less than one year) and (31-50)years was scored 3(100%), 11(100%), respectively.