Author/Authors :
Nazemalhosseini Mojarad, Ehsan shahid beheshti university of medical sciences - School of Medicine - Department of Medical Parasitology and Mycology, تهران, ايران , Nochi, Zahra Islamic Azad University, Sciences and Researches Branch - Department of Basic Science, ايران , Sahebekhtiari, Navid Islamic Azad University, North Tehran Branch - School of Basic Science - Department of Microbiology, ايران , Rostami Nejad, Mohammad Islamic Azad University, Sciences and Researches Branch - Department of Basic Science, ايران , Dabiri, Hossein tehran university of medical sciences tums - School of Medicine - Department of Medical Microbiology, تهران, ايران , Haghighi, Ali shahid beheshti university of medical sciences - School of Medicine - Department of Medical Parasitology and Mycology, تهران, ايران
Abstract :
Aim: The aim of this study was to differentiate Entamoeba dispar from Entamoeba histolytica by PCR directly from fresh stool. Background: Microscopy does not allow for the differentiation of Entamoeba dispar from Entamoeba histolytica. Several PCR-based methods have been described and used successfully for this purpose, but the methods for DNA extraction from stool samples are usually time-consuming and problematic due to inhibitory factors in feces. Patients and methods: From a total of 1700 stool samples collected and examined by microscopy, 22 samples (1.3%)were microscopically positive for the E. histolytica /E. dispar complex. The DNA of these samples was extracted directly from fresh stool and PCR was carried out using two sets of species-specific primers from a short tandem repeat (STR) in the D-A locus. Results: Of these, 21 samples (95.45%) were diagnosed as E. dispar and only one sample (4.55 %) was found to be E. histolytica. In this study, by improving the DNA extraction from fresh stool, we were able to efficiently differentiate E.histolytica and E. dispar. Conclusion: To avoid unnecessary treatment of patients not infected with E. histolytica, the development of effective techniques, such as direct DNA extraction from stool, is recommended.
Keywords :
Entamoeba histolytica , Entamoeba dispar , fresh stool , PCR , locus D , A
