Please use this identifier to cite or link to this item: http://repository.aaup.edu/jspui/handle/123456789/1564
Title: Concurrent molecular characterization of sand flies and Leishmania parasites by amplicon-based next-generation sequencing
Authors: Nasereddin, Abedelmajeed $Other$Palestinian
Ereqat, Suheir$Other$Palestinian
Al-Jawabreh, Amer$AAUP$Palestinian
Taradeh, Mohamad$Other$Palestinian
Abbasi, Ibrahim$Other$Palestinian
Al-Jawabreh, Hanan$Other$Palestinian
Sawalha, Samer$Other$Palestinian
Abdeen, Ziad$Other$Palestinian
Keywords: Amp-NGS
Leishmania
Phlebotomine sand flies
Taxonomy
Issue Date: 22-Jul-2022
Publisher: BMC
Citation: Nasereddin A, Ereqat S, Al-Jawabreh A, Taradeh M, Abbasi I, Al-Jawabreh H, Sawalha S, Abdeen Z. Concurrent molecular characterization of sand flies and Leishmania parasites by amplicon-based next-generation sequencing. Parasit Vectors. 2022 Jul 22;15(1):262. doi: 10.1186/s13071-022-05388-3. PMID: 35869485; PMCID: PMC9308317.
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.
URI: http://repository.aaup.edu/jspui/handle/123456789/1564
ISSN: 10.1186/s13071-022-05388-3
Appears in Collections:Faculty & Staff Scientific Research publications

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