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Online Journal of Veterinary Research©
Volume 22 (4):288-289, 2018.
Frequency of Entamoeba histolytica genotypes in diarrheic patients determined by nested RT-PCR’s.
Russul Wassit Kadhum1, Nawar Jaber Hussein Al–Asadi2, Baraa Abdulsalam Hraiga3
1Department of Biology, College of Science, 2Microbiology, 3College of Medicine, Wasit University, Kut, Iraq.
Kadhum RW, Hussein Al–Asadi NJ, Hraiga BA., Frequency of Entamoeba histolytica genotypes in diarrheic patients determined by nested RT-PCR’s, Onl J Vet Res., 22 (4):288-289, 2018. A nested PCR for detection of Entamoeba histolytica from 50 patients in Wasit, Iraq with diarrhea from stools samples by 2 amplifications of a sub-unit RNA is described. Thirty stools were found to be positive and subjected to another RT-PCR to identify genotypes using specific primers polymorphic for serine rich E. histolytica protein (SREHP). In a 1st run, a conventional PCR amplified a 549-bp fragment of the SREHP gene. In the 2nd run, a nested RT-PCR was done with SYBR Green I. Genotypes were identified by SYBR Green I melting curve analysis. We found 5 different genotypes classified as I to V with frequencies of II 46.67%, I 13.33%, III and ӀV 16.67% and V only 6.7%. Our nested and RT-PCR’s may be an alternative method for rapid screening for amebiasis.
Keywords: Molecular detection; Genotyping; Entamoeba histolytica.