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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical
Society
2020
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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. |
format | Online Article Text |
id | pubmed-7203758 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | American Chemical
Society |
record_format | MEDLINE/PubMed |
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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