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Molecular identification and Phylogenetic Analysis of Enterobius vermicularis isolated from children in Thi-Qar City of Iraq.
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| Author:
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RUAA MAJID KHAZAAL, SALEEM KHTEER AL-HADRAAWY, KHWAM REISSAN HUSSEIN
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| Abstract:
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Background: E. vermicularis is one of the most prevalence helminths in Iraq especially among children because of the children tendency to harbour highest number of intestinal parasitic infestations. The molecular characterization studies of E. vermicularis were limited in Iraq, and molecular diagnosis were associated with some difficulties.
Aim of study: To diagnose E. vermicularis using cox1 and 12S mtDNA, also ITS1 and 18S rRNA genes and determine genetic vareiation using 18S rRNA among samples of enterobiasis infected children,and to sequence the local E. vermicularis isolates and deposits them in global GenBank (NCBI), also to make a comparison between our local isolates and global NCBI records.
Methodology: the current study was conducted in Thi-Qar province south of Iraq for the period from November 2018 to October 2019, and include collection of stool samples to children who had a clinical symptoms for enterobiasis, and diagnosed microscopically (using direct method) as positive to E. vermicularis ova, extraction of genomic DNA was accomplished for all samples, then diagnosis via PCR technique were carried out and analysis of PCR products was performed using agarose gel electrophoresis. The analysis of DNA sequencing data was carried out using Molecular Evolutionary Genetics Analysis version 6 (Mega-6) and multiple sequence alignment analysis of 18S sequences based on Clustal W alignment analysis program.
Results: The results of this study showed that 28 (52.83%) samples were positive of total 53 samples that diagnosed as positive samples by direct wet mount technique, while 25 (47.16%) were negative for the four genes in this study. Agarose gel electrophoresis to PCR products for ITS1 gene was positive at 580bp, cox1 gene was positive at 566 bp, 18S gene was positive at 215 bp, 12S gene was positive at 500 bp. The DNA sequencing analysis for 8 human E. vermicularis local isolates (IQN,No.1 - IQN,No.8 small subunit ribosomal RNA gene) revealed that there were some nucleotide substitution in these local isolates. All these eight local isolates showed 99% homology identity with NCBI-BLAST submitted E. vermicularis isolates, and showed genetic related to NCBI-BLAST E. vermicularis at total genetic changes (0.0020%).
Conclusion: The local isolates that isolated from enterobiasis infected children showed 99% identity homology with NCBI-BLAST submitted E. vermicularis human isolates, and the phylogenetic anaylsis for these eight local isolates showed genetic related to NCBI-BLAST E. vermicularis at total genetic changes (0.0020%).
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| Keyword:
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Enterobius vermicularis, Molecular identification, Phylogenetic Analysis.
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| EOI:
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| DOI:
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https://doi.org/10.31838/ijpr/2020.SP1.293
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| Download:
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