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Designing specific primers for the in-house detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii
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Minoo Shaddel   , Mohammad Rajabloo , Mohammad Foad Heydary , Mohamad Mohsen Homayoni , Javid Imani Azadtalab |
| Department of Parasitology, Faculty of Medicine, AJA University of Medical Sciences, Tehran, Iran. |
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Abstract: (17 Views) |
Introduction: Amoebiasis is caused by the intestinal protozoan parasite Entamoeba histolytica. Accurate differentiation between parasite species is crucial for appropriate clinical management. Nowadays, the use of molecular methods has become one of the important tools in species differentiation. This study aimed to develop a simple, efficient, in-house method that is suitable for laboratories with minimal molecular equipment and serves as a low-cost screening test for the identification of these amoebae.
Methods: A species-specific primer set based on the 18S rRNA gene was designed to differentiate between Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii using bioinformatics tools. In the next step, the efficacy, specificity and sensitivity of these primers were investigated using samples that had been previously identified in studies, including the HM-1:IMSS strain of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii. For this purpose, DNA of the above parasites was first extracted and evaluated by both conventional PCR and Real-time PCR primers.
Results: The primers developed in this study were successful in detecting Entamoeba species and can be used to differentiate Entamoeba species in microscopically positive cases. The primers effectively identified the target species with a detection limit of 10 trophozoites per gram of feces and 2.5 trophozoites per milliliter of water.
Conclusion: According to the results, these primers can be used for rapid detection and also in future studies as probes for the development of more accurate and quantitative diagnostic kits suitable for in vitro diagnostic applications.
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Article number: 03 |
| Keywords: Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, Molecular identification |
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Full-Text [PDF 226 kb]
(10 Downloads)
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Type of Study: Applicable |
Subject:
Special
Received: 2025/12/20 | Accepted: 2026/07/1 | Published: 2026/10/10
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