Selective whole-genome amplification reveals population genetics of Leishmania braziliensis directly from patient skin biopsies.

In Brazil, Leishmania braziliensis is the main causative agent of the neglected tropical disease, cutaneous leishmaniasis (CL). CL presents on a spectrum of disease severity with a high rate of treatment failure. Yet the parasite factors that contribute to disease presentation and treatment outcome...

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Main Authors: Olivia A Pilling, João L Reis-Cunha, Cooper A Grace, Alexander S F Berry, Matthew W Mitchell, Jane A Yu, Clara R Malekshahi, Elise Krespan, Christina K Go, Cláudia Lombana, Yun S Song, Camila F Amorim, Alexsandro S Lago, Lucas P Carvalho, Edgar M Carvalho, Dustin Brisson, Phillip Scott, Daniel C Jeffares, Daniel P Beiting
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2023-03-01
Series:PLoS Pathogens
Online Access:https://doi.org/10.1371/journal.ppat.1011230
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author Olivia A Pilling
João L Reis-Cunha
Cooper A Grace
Alexander S F Berry
Matthew W Mitchell
Jane A Yu
Clara R Malekshahi
Elise Krespan
Christina K Go
Cláudia Lombana
Yun S Song
Camila F Amorim
Alexsandro S Lago
Lucas P Carvalho
Edgar M Carvalho
Dustin Brisson
Phillip Scott
Daniel C Jeffares
Daniel P Beiting
author_facet Olivia A Pilling
João L Reis-Cunha
Cooper A Grace
Alexander S F Berry
Matthew W Mitchell
Jane A Yu
Clara R Malekshahi
Elise Krespan
Christina K Go
Cláudia Lombana
Yun S Song
Camila F Amorim
Alexsandro S Lago
Lucas P Carvalho
Edgar M Carvalho
Dustin Brisson
Phillip Scott
Daniel C Jeffares
Daniel P Beiting
author_sort Olivia A Pilling
collection DOAJ
description In Brazil, Leishmania braziliensis is the main causative agent of the neglected tropical disease, cutaneous leishmaniasis (CL). CL presents on a spectrum of disease severity with a high rate of treatment failure. Yet the parasite factors that contribute to disease presentation and treatment outcome are not well understood, in part because successfully isolating and culturing parasites from patient lesions remains a major technical challenge. Here we describe the development of selective whole genome amplification (SWGA) for Leishmania and show that this method enables culture-independent analysis of parasite genomes obtained directly from primary patient skin samples, allowing us to circumvent artifacts associated with adaptation to culture. We show that SWGA can be applied to multiple Leishmania species residing in different host species, suggesting that this method is broadly useful in both experimental infection models and clinical studies. SWGA carried out directly on skin biopsies collected from patients in Corte de Pedra, Bahia, Brazil, showed extensive genomic diversity. Finally, as a proof-of-concept, we demonstrated that SWGA data can be integrated with published whole genome data from cultured parasite isolates to identify variants unique to specific geographic regions in Brazil where treatment failure rates are known to be high. SWGA provides a relatively simple method to generate Leishmania genomes directly from patient samples, unlocking the potential to link parasite genetics with host clinical phenotypes.
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institution Kabale University
issn 1553-7366
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publishDate 2023-03-01
publisher Public Library of Science (PLoS)
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spelling doaj-art-737717bbee8b47e590ec22cd8859a3a42025-08-20T03:57:59ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742023-03-01193e101123010.1371/journal.ppat.1011230Selective whole-genome amplification reveals population genetics of Leishmania braziliensis directly from patient skin biopsies.Olivia A PillingJoão L Reis-CunhaCooper A GraceAlexander S F BerryMatthew W MitchellJane A YuClara R MalekshahiElise KrespanChristina K GoCláudia LombanaYun S SongCamila F AmorimAlexsandro S LagoLucas P CarvalhoEdgar M CarvalhoDustin BrissonPhillip ScottDaniel C JeffaresDaniel P BeitingIn Brazil, Leishmania braziliensis is the main causative agent of the neglected tropical disease, cutaneous leishmaniasis (CL). CL presents on a spectrum of disease severity with a high rate of treatment failure. Yet the parasite factors that contribute to disease presentation and treatment outcome are not well understood, in part because successfully isolating and culturing parasites from patient lesions remains a major technical challenge. Here we describe the development of selective whole genome amplification (SWGA) for Leishmania and show that this method enables culture-independent analysis of parasite genomes obtained directly from primary patient skin samples, allowing us to circumvent artifacts associated with adaptation to culture. We show that SWGA can be applied to multiple Leishmania species residing in different host species, suggesting that this method is broadly useful in both experimental infection models and clinical studies. SWGA carried out directly on skin biopsies collected from patients in Corte de Pedra, Bahia, Brazil, showed extensive genomic diversity. Finally, as a proof-of-concept, we demonstrated that SWGA data can be integrated with published whole genome data from cultured parasite isolates to identify variants unique to specific geographic regions in Brazil where treatment failure rates are known to be high. SWGA provides a relatively simple method to generate Leishmania genomes directly from patient samples, unlocking the potential to link parasite genetics with host clinical phenotypes.https://doi.org/10.1371/journal.ppat.1011230
spellingShingle Olivia A Pilling
João L Reis-Cunha
Cooper A Grace
Alexander S F Berry
Matthew W Mitchell
Jane A Yu
Clara R Malekshahi
Elise Krespan
Christina K Go
Cláudia Lombana
Yun S Song
Camila F Amorim
Alexsandro S Lago
Lucas P Carvalho
Edgar M Carvalho
Dustin Brisson
Phillip Scott
Daniel C Jeffares
Daniel P Beiting
Selective whole-genome amplification reveals population genetics of Leishmania braziliensis directly from patient skin biopsies.
PLoS Pathogens
title Selective whole-genome amplification reveals population genetics of Leishmania braziliensis directly from patient skin biopsies.
title_full Selective whole-genome amplification reveals population genetics of Leishmania braziliensis directly from patient skin biopsies.
title_fullStr Selective whole-genome amplification reveals population genetics of Leishmania braziliensis directly from patient skin biopsies.
title_full_unstemmed Selective whole-genome amplification reveals population genetics of Leishmania braziliensis directly from patient skin biopsies.
title_short Selective whole-genome amplification reveals population genetics of Leishmania braziliensis directly from patient skin biopsies.
title_sort selective whole genome amplification reveals population genetics of leishmania braziliensis directly from patient skin biopsies
url https://doi.org/10.1371/journal.ppat.1011230
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