Application of Multiplex PCR for Detection and Differentiation of Entamoeba histolytica,Entamoeba dispar and Entamoeba moshkovskii |
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| Authors: | Nozhat ZEBARDAST Ali HAGHIGHI Farshid YEGANEH Seyyed Javad SEYYED TABAEI Mohammad Javad GHARAVI Shirzad FALLAHI Zohreh LASJERDI Nima SALEHI Niloofar TAGHIPOUR Cobra KOHANSAL Farideh NADERI |
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| Institution: | 1.Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran;2.Dept. of Immunology, Shahid Beheshti University of Medical Sciences, Tehran, Iran;3.Dept. of Parasitology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran;4.Dept. of Parasitology and Mycology, Lorestan University of Medical Sciences, Khorramabad, Iran;5.Tamin Ejtemaee Hospital, Ahvaz, Iran |
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| Abstract: | Background
Entamoeba moshkovskii and E. dispar are impossible to differentiate microscopically from the pathogenic species E. histolytica. Multiplex polymerase chain reaction (Multiplex PCR) is a widespread molecular biology technique for amplification of multiple targets in a single PCR experiment.MethodsFor detection and differentiation of the three-microscopy indistinguishable Entamoeba species in human, multiplex PCR assay using different DNA extraction methods was studied. A conserved forward primer was derived from the middle of the small-subunit rRNA gene, and reverse primers were designed from signature sequences specific to each of these three Entamoeba species.ResultsA 166-bp PCR product with E. histolytica DNA, a 580-bp product with E. moshkovskii DNA and a 752-bp product with E. dispar DNA were generated in a single-round and multiplex PCR reaction.ConclusionWe recommend this PCR assay as an accurate, rapid, and effective diagnostic method for the detection and discrimination of these three Entamoeba species in both routine diagnosis of amoebiasis and epidemiological surveys. |
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| Keywords: | Entamoeba hitolytica Entamoeba dispar Entamoeba moshkovskii DNA extraction Multiplex PCR |
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