Concurrent molecular characterization of sand flies and Leishmania parasites by amplicon-based next-generation sequencing.

Autor: Nasereddin A; Biochemistry and Molecular Biology Department, Faculty of Medicine, Al-Quds University, Abu Deis, East Jerusalem, Palestine., Ereqat S; Biochemistry and Molecular Biology Department, Faculty of Medicine, Al-Quds University, Abu Deis, East Jerusalem, Palestine. sereqat@staff.alquds.edu., Al-Jawabreh A; Department of Medical Laboratory Sciences, Faculty of Allied Health Sciences, Arab American University, Jenin, Palestine.; Leishmaniases Research Unit, Jericho, Palestine., Taradeh M; Al-Quds Nutrition and Health Research Institute, Al-Quds University, East Jerusalem, Palestine.; AL-Quds Public Health Society, East Jerusalem, Palestine., Abbasi I; Department of Biology and Biotechnology, Al-Quds University, East Jerusalem, Palestine., Al-Jawabreh H; Leishmaniases Research Unit, Jericho, Palestine.; Al-Quds Nutrition and Health Research Institute, Al-Quds University, East Jerusalem, Palestine.; AL-Quds Public Health Society, East Jerusalem, Palestine., Sawalha S; Ministry of Health, Ramallah, Palestine., Abdeen Z; Al-Quds Nutrition and Health Research Institute, Al-Quds University, East Jerusalem, Palestine.; AL-Quds Public Health Society, East Jerusalem, Palestine.
Jazyk: angličtina
Zdroj: Parasites & vectors [Parasit Vectors] 2022 Jul 22; Vol. 15 (1), pp. 262. Date of Electronic Publication: 2022 Jul 22.
DOI: 10.1186/s13071-022-05388-3
Abstrakt: 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.
(© 2022. The Author(s).)
Databáze: MEDLINE
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