Assessing anti-malarial drug effects ex vivo using the haemozoin detection assay

Detalhes bibliográficos
Autor(a) principal: Rebelo, Maria
Data de Publicação: 2015
Outros Autores: Tempera, Carolina, Fernandes, José F, Grobusch, Martin P, Hanscheid, Thomas
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10451/53177
Resumo: © 2015 Rebelo et al.; licensee BioMed Central. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
id RCAP_caa29944ac034d02282186bdc21c0777
oai_identifier_str oai:repositorio.ul.pt:10451/53177
network_acronym_str RCAP
network_name_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository_id_str 7160
spelling Assessing anti-malarial drug effects ex vivo using the haemozoin detection assayMalariaField trialAnti-malarial sensitivity testingResistanceFlow cytometryHaemozoin© 2015 Rebelo et al.; licensee BioMed Central. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.Background: In vitro sensitivity assays are crucial to detect and monitor drug resistance. Plasmodium falciparum has developed resistance to almost all anti-malarial drugs. Although different in vitro drug assays are available, some of their inherent characteristics limit their application, especially in the field. A recently developed approach based on the flow cytometric detection of haemozoin (Hz) allowed reagent-free monitoring of parasite maturation and detection of drug effects in culture-adapted parasites. In this study, the set-up, performance and usefulness of this novel assay were investigated under field conditions in Gabon. Methods: An existing flow cytometer (Cyflow Blue) was modified on site to detect light depolarization caused by Hz. Blood from malaria patients was incubated for 72 hrs with increasing concentrations of chloroquine, artesunate and artemisinin. The percentage of depolarizing red blood cells (RBC) was used as maturation indicator and measured at 24, 48 and 72 hrs of incubation to determine parasite growth and drug effects. Results: The flow cytometer was easily adapted on site to detect light depolarization caused by Hz. Analysis of ex vivo cultures of parasites, obtained from blood samples of malaria patients, showed four different growth profiles. In 39/46 samples, 50% inhibitory concentrations (IC50) were successfully determined. IC50 values for chloroquine were higher than 200 nM in 70% of the samples, indicating the presence of chloroquine-resistant parasites. For artesunate and artemisinin, IC50 values ranged from 0.9 to 60 nM and from 2.2 nM to 124 nM, respectively, indicating fully sensitive parasites. Conclusion: Flow cytometric detection of Hz allowed the detection of drug effects in blood samples from malaria patients, without using additional reagents or complex protocols. Adjustment of the initial parasitaemia was not required, which greatly simplifies the protocol, although it may lead to different IC50 values. Further investigation of set-up conditions of the Hz assay, as well as future studies in various settings should be performed to further determine the usefulness of this assay as a tool for rapid resistance testing in malaria-endemic countries.This work was supported by the Luso-American Foundation (FLAD-LACR grant: B-A.V-109-09/07). MR acknowledges Fundação para a Ciência e a Tecnologia for doctoral grant (SFRH/BD/84530/2012) and Fundação Calouste Gulbenkian for the Award CAML/Gulbenkian for Travel ACGT fellowship.Springer NatureRepositório da Universidade de LisboaRebelo, MariaTempera, CarolinaFernandes, José FGrobusch, Martin PHanscheid, Thomas2022-05-25T15:42:07Z20152015-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10451/53177engMalar J. 2015 Apr 1;14:14010.1186/s12936-015-0657-81475-2875info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-11-08T16:58:51Zoai:repositorio.ul.pt:10451/53177Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T22:04:08.915400Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Assessing anti-malarial drug effects ex vivo using the haemozoin detection assay
title Assessing anti-malarial drug effects ex vivo using the haemozoin detection assay
spellingShingle Assessing anti-malarial drug effects ex vivo using the haemozoin detection assay
Rebelo, Maria
Malaria
Field trial
Anti-malarial sensitivity testing
Resistance
Flow cytometry
Haemozoin
title_short Assessing anti-malarial drug effects ex vivo using the haemozoin detection assay
title_full Assessing anti-malarial drug effects ex vivo using the haemozoin detection assay
title_fullStr Assessing anti-malarial drug effects ex vivo using the haemozoin detection assay
title_full_unstemmed Assessing anti-malarial drug effects ex vivo using the haemozoin detection assay
title_sort Assessing anti-malarial drug effects ex vivo using the haemozoin detection assay
author Rebelo, Maria
author_facet Rebelo, Maria
Tempera, Carolina
Fernandes, José F
Grobusch, Martin P
Hanscheid, Thomas
author_role author
author2 Tempera, Carolina
Fernandes, José F
Grobusch, Martin P
Hanscheid, Thomas
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Repositório da Universidade de Lisboa
dc.contributor.author.fl_str_mv Rebelo, Maria
Tempera, Carolina
Fernandes, José F
Grobusch, Martin P
Hanscheid, Thomas
dc.subject.por.fl_str_mv Malaria
Field trial
Anti-malarial sensitivity testing
Resistance
Flow cytometry
Haemozoin
topic Malaria
Field trial
Anti-malarial sensitivity testing
Resistance
Flow cytometry
Haemozoin
description © 2015 Rebelo et al.; licensee BioMed Central. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
publishDate 2015
dc.date.none.fl_str_mv 2015
2015-01-01T00:00:00Z
2022-05-25T15:42:07Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10451/53177
url http://hdl.handle.net/10451/53177
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Malar J. 2015 Apr 1;14:140
10.1186/s12936-015-0657-8
1475-2875
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Springer Nature
publisher.none.fl_str_mv Springer Nature
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv
_version_ 1799134592199819264

Registros relacionados