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Dual-Luciferase-Based Fast and Sensitive Detection of Malaria Hypnozoites for the Discovery of Antirelapse Compounds

[Image: see text] Efforts to eradicate Plasmodium vivax malaria are hampered by the presence of hypnozoites, persisting stages in the liver that can reactivate after prolonged periods of time enabling further transmission and causing renewed disease. Large-scale drug screening is needed to identify...

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Autores principales: Voorberg-van der Wel, Annemarie M., Zeeman, Anne-Marie, Nieuwenhuis, Ivonne G., van der Werff, Nicole M., Klooster, Els J., Klop, Onny, Vermaat, Lars C., Kocken, Clemens H. M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2020
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7203758/
https://www.ncbi.nlm.nih.gov/pubmed/32267675
http://dx.doi.org/10.1021/acs.analchem.0c00547
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author Voorberg-van der Wel, Annemarie M.
Zeeman, Anne-Marie
Nieuwenhuis, Ivonne G.
van der Werff, Nicole M.
Klooster, Els J.
Klop, Onny
Vermaat, Lars C.
Kocken, Clemens H. M.
author_facet Voorberg-van der Wel, Annemarie M.
Zeeman, Anne-Marie
Nieuwenhuis, Ivonne G.
van der Werff, Nicole M.
Klooster, Els J.
Klop, Onny
Vermaat, Lars C.
Kocken, Clemens H. M.
author_sort Voorberg-van der Wel, Annemarie M.
collection PubMed
description [Image: see text] Efforts to eradicate Plasmodium vivax malaria are hampered by the presence of hypnozoites, persisting stages in the liver that can reactivate after prolonged periods of time enabling further transmission and causing renewed disease. Large-scale drug screening is needed to identify compounds with antihypnozoite activity, but current platforms rely on time-consuming high-content fluorescence imaging as read-out, limiting assay throughput. We here report an ultrafast and sensitive dual-luciferase-based method to differentiate hypnozoites from liver stage schizonts using a transgenic P. cynomolgi parasite line that contains Nanoluc driven by the constitutive hsp70 promoter, as well as firefly luciferase driven by the schizont-specific lisp2 promoter. The transgenic parasite line showed similar fitness and drug sensitivity profiles of selected compounds to wild type. We demonstrate robust bioluminescence-based detection of hypnozoites in 96-well and 384-well plate formats, setting the stage for implementation in large scale drug screens.
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spelling pubmed-72037582020-05-07 Dual-Luciferase-Based Fast and Sensitive Detection of Malaria Hypnozoites for the Discovery of Antirelapse Compounds Voorberg-van der Wel, Annemarie M. Zeeman, Anne-Marie Nieuwenhuis, Ivonne G. van der Werff, Nicole M. Klooster, Els J. Klop, Onny Vermaat, Lars C. Kocken, Clemens H. M. Anal Chem [Image: see text] Efforts to eradicate Plasmodium vivax malaria are hampered by the presence of hypnozoites, persisting stages in the liver that can reactivate after prolonged periods of time enabling further transmission and causing renewed disease. Large-scale drug screening is needed to identify compounds with antihypnozoite activity, but current platforms rely on time-consuming high-content fluorescence imaging as read-out, limiting assay throughput. We here report an ultrafast and sensitive dual-luciferase-based method to differentiate hypnozoites from liver stage schizonts using a transgenic P. cynomolgi parasite line that contains Nanoluc driven by the constitutive hsp70 promoter, as well as firefly luciferase driven by the schizont-specific lisp2 promoter. The transgenic parasite line showed similar fitness and drug sensitivity profiles of selected compounds to wild type. We demonstrate robust bioluminescence-based detection of hypnozoites in 96-well and 384-well plate formats, setting the stage for implementation in large scale drug screens. American Chemical Society 2020-04-08 2020-05-05 /pmc/articles/PMC7203758/ /pubmed/32267675 http://dx.doi.org/10.1021/acs.analchem.0c00547 Text en Copyright © 2020 American Chemical Society This is an open access article published under a Creative Commons Attribution (CC-BY) License (http://pubs.acs.org/page/policy/authorchoice_ccby_termsofuse.html) , which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.
spellingShingle Voorberg-van der Wel, Annemarie M.
Zeeman, Anne-Marie
Nieuwenhuis, Ivonne G.
van der Werff, Nicole M.
Klooster, Els J.
Klop, Onny
Vermaat, Lars C.
Kocken, Clemens H. M.
Dual-Luciferase-Based Fast and Sensitive Detection of Malaria Hypnozoites for the Discovery of Antirelapse Compounds
title Dual-Luciferase-Based Fast and Sensitive Detection of Malaria Hypnozoites for the Discovery of Antirelapse Compounds
title_full Dual-Luciferase-Based Fast and Sensitive Detection of Malaria Hypnozoites for the Discovery of Antirelapse Compounds
title_fullStr Dual-Luciferase-Based Fast and Sensitive Detection of Malaria Hypnozoites for the Discovery of Antirelapse Compounds
title_full_unstemmed Dual-Luciferase-Based Fast and Sensitive Detection of Malaria Hypnozoites for the Discovery of Antirelapse Compounds
title_short Dual-Luciferase-Based Fast and Sensitive Detection of Malaria Hypnozoites for the Discovery of Antirelapse Compounds
title_sort dual-luciferase-based fast and sensitive detection of malaria hypnozoites for the discovery of antirelapse compounds
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7203758/
https://www.ncbi.nlm.nih.gov/pubmed/32267675
http://dx.doi.org/10.1021/acs.analchem.0c00547
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