Acanthamoeba castellanii: a new high-throughput method for drug screening in vitro

Despite significant public health impact, there is no specific antiprotozoal therapy for prevention and treatment of Acanthamoeba castellanii infection. There is a need for new and efficient anti-Acanthamoeba drugs that are less toxic and can reduce treatment duration and frequency of administration...

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Main Authors: Ortega-Rivas, Antonio, Padrón, José M., Valladares, Basilio, Elsheikha, Hany M.
Format: Article
Published: Elsevier 2016
Subjects:
Online Access:https://eprints.nottingham.ac.uk/44052/
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author Ortega-Rivas, Antonio
Padrón, José M.
Valladares, Basilio
Elsheikha, Hany M.
author_facet Ortega-Rivas, Antonio
Padrón, José M.
Valladares, Basilio
Elsheikha, Hany M.
author_sort Ortega-Rivas, Antonio
building Nottingham Research Data Repository
collection Online Access
description Despite significant public health impact, there is no specific antiprotozoal therapy for prevention and treatment of Acanthamoeba castellanii infection. There is a need for new and efficient anti-Acanthamoeba drugs that are less toxic and can reduce treatment duration and frequency of administration. In this context a new, rapid and sensitive assay is required for high-throughput activity testing and screening of new therapeutic compounds. A colorimetric assay based on sulforhodamine B (SRB) staining has been developed for anti-Acanthamoeba drug susceptibility testing and adapted to a 96-well microtiter plate format. Under these conditions chlorhexidine was tested to validate the assay using two clinical strains of A. castellanii (Neff strain, T4 genotype [IC50 4.68±0.6 _M] and T3 genotype [IC50 5.69±0.9 _M]). These results were in good agreement with those obtained by the conventional Alamar Blue assay, OCR cytotoxicity assay and manual cell counting method. Our new assay offers an inexpensive and reliable method, which complements current assays by enhancing high-throughput anti-Acanthamoeba drug screening capabilities.
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spelling nottingham-440522020-05-04T18:18:56Z https://eprints.nottingham.ac.uk/44052/ Acanthamoeba castellanii: a new high-throughput method for drug screening in vitro Ortega-Rivas, Antonio Padrón, José M. Valladares, Basilio Elsheikha, Hany M. Despite significant public health impact, there is no specific antiprotozoal therapy for prevention and treatment of Acanthamoeba castellanii infection. There is a need for new and efficient anti-Acanthamoeba drugs that are less toxic and can reduce treatment duration and frequency of administration. In this context a new, rapid and sensitive assay is required for high-throughput activity testing and screening of new therapeutic compounds. A colorimetric assay based on sulforhodamine B (SRB) staining has been developed for anti-Acanthamoeba drug susceptibility testing and adapted to a 96-well microtiter plate format. Under these conditions chlorhexidine was tested to validate the assay using two clinical strains of A. castellanii (Neff strain, T4 genotype [IC50 4.68±0.6 _M] and T3 genotype [IC50 5.69±0.9 _M]). These results were in good agreement with those obtained by the conventional Alamar Blue assay, OCR cytotoxicity assay and manual cell counting method. Our new assay offers an inexpensive and reliable method, which complements current assays by enhancing high-throughput anti-Acanthamoeba drug screening capabilities. Elsevier 2016-12-01 Article PeerReviewed Ortega-Rivas, Antonio, Padrón, José M., Valladares, Basilio and Elsheikha, Hany M. (2016) Acanthamoeba castellanii: a new high-throughput method for drug screening in vitro. Acta Tropica, 164 . pp. 95-99. ISSN 0001-706X Acanthamoeba Colorimetric assay Drug screening Sulforhodamine B http://www.sciencedirect.com/science/article/pii/S0001706X16304533?via%3Dihub doi:10.1016/j.actatropica.2016.09.006 doi:10.1016/j.actatropica.2016.09.006
spellingShingle Acanthamoeba
Colorimetric assay
Drug screening
Sulforhodamine B
Ortega-Rivas, Antonio
Padrón, José M.
Valladares, Basilio
Elsheikha, Hany M.
Acanthamoeba castellanii: a new high-throughput method for drug screening in vitro
title Acanthamoeba castellanii: a new high-throughput method for drug screening in vitro
title_full Acanthamoeba castellanii: a new high-throughput method for drug screening in vitro
title_fullStr Acanthamoeba castellanii: a new high-throughput method for drug screening in vitro
title_full_unstemmed Acanthamoeba castellanii: a new high-throughput method for drug screening in vitro
title_short Acanthamoeba castellanii: a new high-throughput method for drug screening in vitro
title_sort acanthamoeba castellanii: a new high-throughput method for drug screening in vitro
topic Acanthamoeba
Colorimetric assay
Drug screening
Sulforhodamine B
url https://eprints.nottingham.ac.uk/44052/
https://eprints.nottingham.ac.uk/44052/
https://eprints.nottingham.ac.uk/44052/