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The present investigational study was intentionally aimed to detect of E.moshkoviskii in stool as the first study and new record in Iraq by using polymerase chain reaction technique. Additionally understand the prevalence of these three parasites (E.moshkoviskii, E.dispar, and E.histolytica) in the human population. Stool sampling of 190 specimens was performed from clinically recognized patients suffering diarrhea with an inflammatory feature from Al-Diwaniya Teaching Hospital, Maternity and Children Hospital, Afak general Hospital and some Hospitals in the province of Iraq. All samples undergo full history was distributed according to name, age, gender, address, bloody diarrhea and clinical symptoms. General stool examination was done to samples within 30 minutes. The samples were divided into two sterile containers the first one grown in media and the second two were kept frozen at -20 Ċ (deep freezing) for DNA extraction used for PCR. From extracted DNA was added 5µl in PCR tube (master mix) with 1.5 µl forward primer and 1.5µl reverse primer, then 12µl from nuclease-free water, all volume was completed to 20 µl. One hundred ninety stool samples were collected and examined by general stool examination positive for Entamoeba spp The present study showed the high prevalence of amoebiasis in rural area more than urban area, the result revealed that the infected patients in rural area 119 (62.6%) while in urban area 71 (37.3%) The present study revealed the high percentage (25.2%) of infection with gastrointestinal symptoms under the age of 14 years old and above the age of 45 years. In the present study was showed the PCR product was detected in 182 (96 %) samples, and 8 (4 %) were detected as negative utilizing PCR test. The present study concluded that presence of three-identical species of Entamoeba utilizing PCR to target stool-based DNA of patients expressing gastrointestinal symptoms (E.moshkoviskii, E. dispar, and E.histolytica) could be grown in modified Lock's – egg slant medium . The single round PCR described in this study is a specific and sensitive method for distinguishing between those species.
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