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J Anal Res Clin Med. 2014;2(1): 25-29.
doi: 10.5681/jarcm.2014.004
  Abstract View: 1365
  PDF Download: 1096

Original Research

Differential detection of Entamoeba histolytica from Entamoeba dispar by parasitological and nested multiplex polymerase chain reaction methods

esmaiel fallah 1, Abbas Shahbazi 2*, majid yazdanjoii 3, Bahman Rahimi-Esboei 4

1 Professor, Department of Parasitology and Mycology, School of Public Health, Tabriz University of Medical Sciences, Tabriz, Iran
2 Associate Professor, Department of Medical Parasitology, Tabriz Research Center of Infectious and Tropical Diseases, Tabriz University
3 Department of Parasitology and Mycology, School of Public Health, Tabriz University of Medical Sciences, Tabriz, Iran
4 PhD Student, Department of Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
*Corresponding Author: Email: shabazy45@yahoo.com

Abstract

Introduction: Amebiasis is an intestinal illness caused by a one-celled parasite (amoeba) called Entamoeba (E) histolytica. E histolytica and E dispar are morphologicallyundistinguishable but have genetic and functional differences. E. histolytica is invasive andcause amoebiasis, but E dispar cause an asymptomatic colonization which does not need to bemedically treated. We have performed a nested multiplex Polymerase Chain Reaction (PCR)targeting small subunit rRNA (Ribosomal ribonucleic acid) gene for differential detection of Ehistolytica and E dispar directly from stool samples. Methods: All the fecal samples collected without preservation and were screened for amebiccells by parasitological methods. Fecal samples that containing amebic cells were stored at  -20ºC until DNA extraction. DNA extraction was down by using a DNA extraction kit. Thegenus specific primers were designed using nucleotide sequences of 18S-rRNA gene ofEntamoeba. Results: Thirty one (4.28%) stool samples out of 724 samples were positive for E histolytica/E dispar. The nested multiplex PCR illustrated that the size of diagnostic fragments of PCR products was obviously different for two Entamoeba species, the specific product size for Ehistolytica and E dispar was 439 and 174 bp. The nested multiplex PCR was positive in 25 outof 31 stool specimens that 17 (54.8%) samples were positive for E dispar and 8 (25.8%)samples were positive for E histolytica. Conclusion: Nested multiplex PCR was useful for the specific detection of E histolytica and Edispar in stool samples. In current study we detected that E dispar was more prevalent in our study area. 
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Submitted: 20 Nov 2013
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