DIAGNOSIS OF ENTAMOEBA HISTOLYTICA/DISPAR USING CONVENTIONAL AND MOLECULAR TECHNIQUES
Jumana A. Ahmed1, Waheed A. Mousa2, Eman Y. Shoeib1, Nihal A.Hanafy1, Rasha A. Kazamel1
Medical Parasitology Department, Faculty of Medicine, Cairo University1 and Parasitology department, Faculty of Veterinary Medicine, Cairo University2
Amoebiasis is the third leading parasitic cause of death in humans after malaria and schistosomiasis. It has a worldwide distribution. Since E. dispar is morphologically identical to E. histolytica; differential identification of E. histolytica and/or E. dispar is important for epidemiological studies and treatment of amoebiasis. The present study aimed at diagnosis of Entamoeba complex using conventional techniques; species specific detection and differentiation of E.histolytica/dispar DNA in stool samples by molecular techniques. A total of 490 stool samples were collected. All samples were subjected to direct stool examination, ether sedimentation concentration technique. Nested multiplex PCR was applied to the samples proved to be positive for Entamoeba. complex by microscopic examination for species specific detection and differentiation of E.histolytica/dispar DNA in stool samples. Entamoeba complex was detected by direct microscopic examinationin in 39 stool sample which represent 7.9% of the studied population. Infection with Entamoeba complex was found more common in males (53.8%) than females(46.1%). It was also more common in adults (56.4%) than children (43.5%). When nested multiplex PCR was applied to the samples proved to be positive by microscopic examination, all of these samples proved to belong to the Entamoeba species; the most common species found was E.histolytica (84.6%) followed by E.dispar (25.6%) then other Entamoeba species (5.1%). The present study highlighted the effectiveness of Nested multiplex PCR as a highly specific and sensitive technique for species specific detection and differentiation of E.histolytica/dispar DNA in stool samples.
June 2013