Detection and differentiation of Entamoeba histolytica and Entamoeba dispar in clinical samples through PCR-denaturing gradient gel electrophoresis

Abstract

Amebiasis is one of the twenty major causes of disease in Mexico; however, the diagnosis is difficult due to limitations of conventional microscopy-based techniques. In this study, we analyzed stool samples using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) to differentiate between Entamoeba histolytica (pathogenic) and E. dispar (non-pathogenic). The target for the PCR amplification was a small region (228 bp) of the adh112 gene selected to increase the sensitivity of the test. The study involved 62 stool samples that were collected from individuals with complaints of gastrointestinal discomfort. Of the 62 samples, 10 (16.1%) were positive for E. histolytica while 52 (83.9%) were negative. No sample was positive for E. dispar. These results were validated by nested PCR-RFLP (restriction fragment length polymorphism) and suggest that PCR-DGGE is a promising tool to differentiate among Entamoeba infections, contributing to determine the specific treatment for patients infected with E. histolytica, and therefore, avoiding unnecessary treatment of patients infected with the non-pathogenic E. dispar.

The epidemiology of amoebiasis in Thi-Qar Province, Iraq (2015-2020): differentiation of Entamoeba histolytica and Entamoeba dispar using nested and real-time polymerase chain reaction

“…In the small intestine, trophozoites excyst to develop and colonize the colonic region, and then adhere to the mucosal layer of the large intestines. E. histolytica causes intestinal and extra-intestinal infections [ 11 ]. In addition, some trophozoites may be excreted in stool outside the human host, but are not able to survive.…”

A Prevalence study of Entamoeba spp. in Basrah Province using Different Detection Methods

“…Secondly differences in the number of parasites in stool samples. Finally, the fixative and preservative agents used in the study may interfere with cell components resulting in effecting DNA (4).…”

“…The main laboratory diagnosis of Entamoeba spp. in human stool is microscopic examination but it is not possible to distinguish between these types depending on it (4).…”

“CLUPS”: A New Culture Medium for the Axenic Growth ofEntamoeba histolytica

“…Ongoing improvement of molecular diagnostic tools facilitates a better differentiation and identification of species [ 5 ]. The incorporation of new diagnostic techniques confirms that the protozoan pathogen Entamoeba histolytica remains a major cause of gastrointestinal disease in Mexico and other parts of the world [ 6 , 7 ]. PCR-based studies reveal that E. histolytica is present in 16-20% of people with a gastrointestinal disorder [ 6 , 7 ].…”

“…The incorporation of new diagnostic techniques confirms that the protozoan pathogen Entamoeba histolytica remains a major cause of gastrointestinal disease in Mexico and other parts of the world [ 6 , 7 ]. PCR-based studies reveal that E. histolytica is present in 16-20% of people with a gastrointestinal disorder [ 6 , 7 ]. Better culture media could facilitate the in vitro study of microorganisms such as E. histolytica .…”