Development and validation of a quantitative, high-throughput, fluorescent-based bioassay to detect schistosoma viability.
Schistosomiasis, caused by infection with the blood fluke Schistosoma, is responsible for greater than 200,000 human deaths per annum. Objective high-throughput screens for detecting novel anti-schistosomal targets will drive 'genome to drug' lead translational science at an unprecedented...
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ftdoajarticles:oai:doaj.org/article:3db52365d6a24519a60d2a3502abbd97 2023-05-15T15:12:45+02:00 Development and validation of a quantitative, high-throughput, fluorescent-based bioassay to detect schistosoma viability. Emily Peak Iain W Chalmers Karl F Hoffmann 2010-07-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0000759 https://doaj.org/article/3db52365d6a24519a60d2a3502abbd97 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC2910722?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0000759 https://doaj.org/article/3db52365d6a24519a60d2a3502abbd97 PLoS Neglected Tropical Diseases, Vol 4, Iss 7, p e759 (2010) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2010 ftdoajarticles https://doi.org/10.1371/journal.pntd.0000759 2022-12-31T11:42:42Z Schistosomiasis, caused by infection with the blood fluke Schistosoma, is responsible for greater than 200,000 human deaths per annum. Objective high-throughput screens for detecting novel anti-schistosomal targets will drive 'genome to drug' lead translational science at an unprecedented rate. Current methods for detecting schistosome viability rely on qualitative microscopic criteria, which require an understanding of parasite morphology, and most importantly, must be subjectively interpreted. These limitations, in the current state of the art, have significantly impeded progress into whole schistosome screening for next generation chemotherapies.We present here a microtiter plate-based method for reproducibly detecting schistosomula viability that takes advantage of the differential uptake of fluorophores (propidium iodide and fluorescein diacetate) by living organisms. We validate this high-throughput system in detecting schistosomula viability using auranofin (a known inhibitor of thioredoxin glutathione reductase), praziquantel and a range of small compounds with previously-described (gambogic acid, sodium salinomycin, ethinyl estradiol, fluoxetidine hydrochloride, miconazole nitrate, chlorpromazine hydrochloride, amphotericin b, niclosamide) or suggested (bepridil, ciclopirox, rescinnamine, flucytosine, vinblastine and carbidopa) anti-schistosomal activities. This developed method is sensitive (200 schistosomula/well can be assayed), relevant to industrial (384-well microtiter plate compatibility) and academic (96-well microtiter plate compatibility) settings, translatable to functional genomics screens and drug assays, does not require a priori knowledge of schistosome biology and is quantitative.The wide-scale application of this fluorescence-based bioassay will greatly accelerate the objective identification of novel therapeutic lead targets/compounds to combat schistosomiasis. Adapting this bioassay for use with other parasitic worm species further offers an opportunity for great strides to be made ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLoS Neglected Tropical Diseases 4 7 e759 |
institution |
Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Emily Peak Iain W Chalmers Karl F Hoffmann Development and validation of a quantitative, high-throughput, fluorescent-based bioassay to detect schistosoma viability. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Schistosomiasis, caused by infection with the blood fluke Schistosoma, is responsible for greater than 200,000 human deaths per annum. Objective high-throughput screens for detecting novel anti-schistosomal targets will drive 'genome to drug' lead translational science at an unprecedented rate. Current methods for detecting schistosome viability rely on qualitative microscopic criteria, which require an understanding of parasite morphology, and most importantly, must be subjectively interpreted. These limitations, in the current state of the art, have significantly impeded progress into whole schistosome screening for next generation chemotherapies.We present here a microtiter plate-based method for reproducibly detecting schistosomula viability that takes advantage of the differential uptake of fluorophores (propidium iodide and fluorescein diacetate) by living organisms. We validate this high-throughput system in detecting schistosomula viability using auranofin (a known inhibitor of thioredoxin glutathione reductase), praziquantel and a range of small compounds with previously-described (gambogic acid, sodium salinomycin, ethinyl estradiol, fluoxetidine hydrochloride, miconazole nitrate, chlorpromazine hydrochloride, amphotericin b, niclosamide) or suggested (bepridil, ciclopirox, rescinnamine, flucytosine, vinblastine and carbidopa) anti-schistosomal activities. This developed method is sensitive (200 schistosomula/well can be assayed), relevant to industrial (384-well microtiter plate compatibility) and academic (96-well microtiter plate compatibility) settings, translatable to functional genomics screens and drug assays, does not require a priori knowledge of schistosome biology and is quantitative.The wide-scale application of this fluorescence-based bioassay will greatly accelerate the objective identification of novel therapeutic lead targets/compounds to combat schistosomiasis. Adapting this bioassay for use with other parasitic worm species further offers an opportunity for great strides to be made ... |
format |
Article in Journal/Newspaper |
author |
Emily Peak Iain W Chalmers Karl F Hoffmann |
author_facet |
Emily Peak Iain W Chalmers Karl F Hoffmann |
author_sort |
Emily Peak |
title |
Development and validation of a quantitative, high-throughput, fluorescent-based bioassay to detect schistosoma viability. |
title_short |
Development and validation of a quantitative, high-throughput, fluorescent-based bioassay to detect schistosoma viability. |
title_full |
Development and validation of a quantitative, high-throughput, fluorescent-based bioassay to detect schistosoma viability. |
title_fullStr |
Development and validation of a quantitative, high-throughput, fluorescent-based bioassay to detect schistosoma viability. |
title_full_unstemmed |
Development and validation of a quantitative, high-throughput, fluorescent-based bioassay to detect schistosoma viability. |
title_sort |
development and validation of a quantitative, high-throughput, fluorescent-based bioassay to detect schistosoma viability. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2010 |
url |
https://doi.org/10.1371/journal.pntd.0000759 https://doaj.org/article/3db52365d6a24519a60d2a3502abbd97 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 4, Iss 7, p e759 (2010) |
op_relation |
http://europepmc.org/articles/PMC2910722?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0000759 https://doaj.org/article/3db52365d6a24519a60d2a3502abbd97 |
op_doi |
https://doi.org/10.1371/journal.pntd.0000759 |
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PLoS Neglected Tropical Diseases |
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4 |
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7 |
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e759 |
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