Concurrent Molecular Characterization of Sand Flies and Leishmania Parasites by Amplicon-based Next-generation Sequencing

Overview

Journal Parasit Vectors

Publisher Biomed Central

Specialties Parasitology
Tropical Medicine

Date 2022 Jul 22

PMID 35869485

Authors

Abedelmajeed Nasereddin

Suheir Ereqat

Amer Al-Jawabreh

Mohamad Taradeh

Ibrahim Abbasi

Hanan Al-Jawabreh

Samer Sawalha

Ziad Abdeen

Affiliations

Soon will be listed here.

Abstract

Background: Phlebotomine sand flies are vectors of Leishmania parasites, which are the causative agents of leishmaniasis. Herein, we developed an amplicon-based next-generation sequencing (Amp-NGS) to characterize sand flies and Leishmania parasites simultaneously targeting partial fragments of 18S rDNA and ITS1 genes, respectively.

Methods: Our assay was optimized using reference sand fly (n = 8) and Leishmania spp. (n = 9) samples and validated using wild-caught sand flies from Palestine. The assay was highly specific, and all DNA references were successfully identified to the species level.

Results: Among the wild-caught sand flies (n = 187), Phlebotomus spp. represented 95% of the collected samples (177/187), including Ph. sergenti (147/187, 79%), Ph. papatasi (19/187, 10.2%), Ph. perfiliewi (3/187, 1.6%), Ph. tobbi (2/187, 1.2%) and Ph. syriacus (6/187, 3.2%). Sergentomyia spp. represented only 5% (10/187) of the collected samples and included S. dentata (n = 6), S. fallax (n = 2), S. schwetzi (n = 1) and S. ghesquiere (n = 1). The study observed strong positive correlation between sand fly identification results of the Amp-NGS and morphological identification method (r = 0.84, df = 185, P < 0.001). Some discrepancies between the two methods in the identification of closely related species (i.e. Ph. perfiliewi, Ph. tobbi and Ph. syriacus) were observed. Leishmania DNA was detected and identified as L. tropica in 14 samples (14/187, 7.5%).

Conclusions: Our assay was sensitive to detect (limit of detection was 0.0016 ng/reaction) and identify Leishmania DNA in sand flies, thus representing a new tool for studying sand flies and their associated Leishmania parasites in endemic areas.

Citing Articles

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PMID: 40034868 PMC: 11873808. DOI: 10.3389/fpara.2025.1547701.

Genetic diversity and haplotype analysis of Leishmania tropica identified in sand fly vectors of the genera Phlebotomus and Sergentomyia using next-generation sequencing technology.

Al-Jawabreh A, Ereqat S, Al-Jawabreh H, Dumaidi K, Nasereddin A Parasitol Res. 2023; 122(6):1351-1360.

PMID: 37036520 DOI: 10.1007/s00436-023-07835-1.

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