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

Ortega-Rivas, Antonio; Padrón, José M.; Valladares, Basilio; Elsheikha, Hany M.

Authors

Antonio Ortega-Rivas

José M. Padrón

Basilio Valladares



Abstract

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.

Citation

Ortega-Rivas, A., PadrĂ³n, J. M., Valladares, B., & Elsheikha, H. M. (2016). Acanthamoeba castellanii : A new high-throughput method for drug screening in vitro. Acta Tropica, 164, 95-99. https://doi.org/10.1016/j.actatropica.2016.09.006

Journal Article Type Article
Acceptance Date Sep 4, 2016
Online Publication Date Sep 5, 2016
Publication Date 2016-12
Deposit Date Jul 7, 2017
Publicly Available Date Jul 7, 2017
Journal Acta Tropica
Print ISSN 0001-706X
Electronic ISSN 1873-6254
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 164
Pages 95-99
DOI https://doi.org/10.1016/j.actatropica.2016.09.006
Keywords Acanthamoeba, Colorimetric assay, Drug screening, Sulforhodamine B
Public URL http://eprints.nottingham.ac.uk/id/eprint/44052
Publisher URL http://www.sciencedirect.com/science/article/pii/S0001706X16304533?via%3Dihub
Copyright Statement Copyright information regarding this work can be found at the following address: http://eprints.nottingh.../end_user_agreement.pdf

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Copyright Statement
Copyright information regarding this work can be found at the following address: http://eprints.nottingham.ac.uk/end_user_agreement.pdf





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