Differential detection of Entamoeba species in stool samples collected from children in District Swat, Khyber Pakhtunkhwa Pakistan

Abstract

29Background: Amoebiasis is an intestinal disease caused by enteric protozoan called 30 Entamoeba histolytica belongs to the Genus Entamoeba. The main reason of infection is the 31 contamination of food and water due to the poor sanitation. Among Entamoeba species, 32 Entamoeba histolytica is highly pathogenic while the other species are non-pathogenic and 33 needs no medical treatment. 34Methodology: A total of 400 stool samples were collected from different areas of 35 district Swat and were processed for screening of amoebic cells. Microscopically identified 36 samples containing amoebic cells were stored at -20 o C till DNA extraction. Extracted DNA 37 was used in a PCR reaction with specific reference primers to amplify the target DNA. 38 Results: Out of all 400 stool samples 111 (27.7%) were found positive through 39 microscopy while PCR reaction confirmed 80 out of microscope positive samples. Among 80 40 PCR positive samples, the infection with Entamoeba dispar was most common (57.5%) 41followed by E. histolytica (47.5%) and Entamoeba moshkovskii (20%). The positive cases for 42 mono-infection of E. dispar were 33 (41.25%), followed by E.histolytica 25 (31.25%) and E. 43 moshkovskii 7 (8.75%). The co-infection of E. histolytica with E. dispar and E. moshkovskii 44 was 6 (7.5%) and 2 (2.5%), respectively. Similarly the co-infection of Entamoeba dispar with 45 Entamoeba moshkovskii was also 2 (2.5%) while 5 (6.25%) samples were observed with 46 mixed infection of E. histolytica, E. dispar and E. moshkovskii. 47 Significance of the study: The aim of the study was to detect and differentiate the 48 E. histolytica, Entamoeba dispar and Entamoeba moshkovskii using conventional microscopy 49 and polymerase chain reaction. The results suggested that the use of PCR is necessary to 50 differentiate E. histolytica from E. dispar and E. moshkovskii and therefore, to avoid 51 unnecessary treatment the present study recommend the use of PCR for the routine diagnosis 52 of amoebiasis in the study area. It is also suggested that further studies from this area may 53 also facilitate the understanding of genetic diversity of these pathogens. 54 55 56 57 58 59 60 61 62 63 65Entamoeba histolytica is an enteric protozoan belongs to genus Entamoeba which is 66 the causative agent of amoebiasis [1]. The Genus Entamoeba comprises of several species 67 among them six reside in human intestinal tract including E. histolytica, E. dispar, E. 68 moshkovskii, E. coli, E. poleki and E. hartmanni [2]. There are two forms Entamoeba, the 69 trophozoite which has short life span and are mobile that can invade the different organ 70 systems, while the cysts are long surviving form of Entamoeba that colonize the patient. The 71 cyst consists of resistant walls which protect these species from desiccation in external 72 environment [3]. In external environment the cysts can survive for several days, weeks or 73 month mostly in damp conditions and are responsible for transmission of infection [4].74 The transmission of ...

molecular identification of entamoeba Dispar and entamoeba Moshkovskii by using nested multiplex PCR in symptomatic indivisuals

“…Our presents study were nearly result for Ngui et al, (2012) recorded 69.3% (52/75) by nested multiplex PCR in Malaysia . In addition, Khan et al, (2019) use PCR reaction to confirmed 72.07% (80/111) out of microscope positive samples real for the parasites in fecal specimens .…”

Molecular identification of Entamoeba spp. in humans and cattle in Baghdad, Iraq

“…Khan et al . [ 31 ] reported a different prevalence, E. dispar had higher prevalence (57.5%) than E. histolytica (47.5%). Another study reported a prevalence of 10% with E. dispar and 5% with E. histolytica in humans [ 11 ].…”

“…Contracting E. histolytica infections can be increased by keeping pets due to the parasite’s prevalence in animals and its spread through contamination. Therefore, it is proposed that more research should be conducted to identify and determine the genetic diversity of these parasites, as well as to determine the true pathogenicity and risk factors associated with Entamoeba species [ 31 ].…”