Folia Parasitologica 71:014 (2024) | DOI: 10.14411/fp.2024.014

Use of the EmsB microsatellite-based next generation sequencing for genotyping of Echinococcus granulosus sensu lato in hydatid cyst tissue samples from animals and humans

Suheir Ereqat ORCID...1, Amer Al-Jawabreh ORCID...2, 3, *, Hanan Al-Jawabreh ORCID...1, 3, Abedelmajeed Nasereddin ORCID...1, 4
1 Biochemistry and Molecular Biology Department, Faculty of Medicine, Al-Quds University, Abu Deis, Jerusalem, Palestine;
2 Department of Medical Laboratory Sciences, Faculty of Allied Health Sciences, Arab American University, Jenin, Palestine;
3 Leishmaniases Research Unit, Jericho, Palestine;
4 Al-Quds Bard College Al-Quds University Jerusalem, Palestine *Address for correspondence: Al-Quds str., Jericho, Palestine, P5840227. E-mail: islahjr@yahoo.com; amer.aljawabreh@aaup.edu

Echinococcus granulosus (Batsch, 1786), a cestode of the Teniidae family, causes human cystic echinococcosis (CE) also known as hydatid disease. Echinococcus granulosus sensu lato includes the G1, G3, G4, G5, G6/7 and G8/10 genotypes which are known to cause human CE. This study aimed to differentiate genotypes of E. granulosus s.l. complex by employing EmsB, a tandemly repeated multilocus microsatellite, using next-generation sequencing (MIC-NGS). Human and animal histopathology-confirmed hydatid cyst tissue samples and reference DNA samples of E. granulosus G1, G3, G4, G5, G6/7 and G10 underwent MIC-NGS assay with custom primers amplifying a 151 bp EmsB DNA fragment. NGS data were analysed using online Galaxy analysis pipeline, a phylogenetic tree was constructed by MEGA software, and haplotype networking was performed with PopArt 1.7. All sixty samples (49 from animals and 11 from humans) included were successfully identified and genotyped with a 100 % success rate. The study showed improved discrimination power to distinguish all study samples including closely related E. granulosus s.s. genotypes G1-G3. The maximum likelihood tree reaffirmed the monophyly of E. granulosus s.l. The median-joining haplotype networking revealed 12 distinct haplotypes. In conclusion, MIC-NGS assay was shown to be sensitive, specific and simple to apply to clinical samples offering a powerful discriminatory tool for the genotyping of E. granulosus s.l.

Keywords: Echinococcus granulosus sensu stricto, echinococcosis, EmsB, genotyping, microsatellite, next-generation sequencing

Received: April 20, 2024; Revised: June 26, 2024; Accepted: July 12, 2024; Published online: August 5, 2024  Show citation

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Ereqat, S., Al-Jawabreh, A., Al-Jawabreh, H., & Nasereddin, A. (2024). Use of the EmsB microsatellite-based next generation sequencing for genotyping of Echinococcus granulosus sensu lato in hydatid cyst tissue samples from animals and humans. Folia Parasitologica, 71, Article 2024.014. https://doi.org/10.14411/fp.2024.014
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