PrestoBlue® and AlamarBlue® are equally useful as agents to determine the viability of Acanthamoeba trophozoites.
Acanthamoeba is an opportunistic pathogen which is the causal agent of several human infections such as Granulomatous Amoebic Encephalitis, Acanthamoeba keratitis and other disseminated infections. Furthermore, current therapeutic measures against Acanthamoeba infections are arduous, and show limited efficacy against the cyst stage of Acanthamoeba. There is a pressing need to search and evaluate new therapeutic agents against these protozoa. Our approach for evaluating possible new drugs is an initial in vitro screening assay based on general metabolic activity of the cells. In this study we compare two agents, AlamarBlue® and PrestoBlue® for this initial screen. Both reagents can be used to indicate metabolism by changes in their absorbance or fluorescence. The assay is carried out in a 96-well plate format and fluorescence can be measured after an inoculation period of as little as 10 min, but more typically 96 h. This to the best of our knowledge this is the first time that both compounds are directly compared using absorbance and fluorescence measurement. We conclude that for the specific case of Acanthamoeba both agents AlamarBlue® and PrestoBlue® are equally useful to determine cell viability.
University Institute of Tropical Diseases and Public Health of the Canary Islands, University of La Laguna, Avda. Astrofísico Fco. Sánchez, S/N, 38203 La Laguna, Tenerife, Canary Islands, Spain; Centre for Integrative Physiology, School of Biomedical Sciences, University of Edinburgh, Edinburgh, Scotland, United Kingdom. Electronic address: cmmartin@ull.es.
Carmen M
CM
Martín-Navarro
University Institute of Tropical Diseases and Public Health of the Canary Islands, University of La Laguna, Avda. Astrofísico Fco. Sánchez, S/N, 38203 La Laguna, Tenerife, Canary Islands, Spain.
Atteneri
A
López-Arencibia
Laboratoire Matériaux-Molécules et Applications, IPEST, B.P. 51, 2070 La Marsa, Tunisia.
Ines
I
Sifaoui
University Institute of Tropical Diseases and Public Health of the Canary Islands, University of La Laguna, Avda. Astrofísico Fco. Sánchez, S/N, 38203 La Laguna, Tenerife, Canary Islands, Spain.
María
M
Reyes-Batlle
University Institute of Tropical Diseases and Public Health of the Canary Islands, University of La Laguna, Avda. Astrofísico Fco. Sánchez, S/N, 38203 La Laguna, Tenerife, Canary Islands, Spain.
Alfonso M
AM
Cabello-Vílchez
Centre for Integrative Physiology, School of Biomedical Sciences, University of Edinburgh, Edinburgh, Scotland, United Kingdom.
Sutherland
S
Maciver
University Institute of Tropical Diseases and Public Health of the Canary Islands, University of La Laguna, Avda. Astrofísico Fco. Sánchez, S/N, 38203 La Laguna, Tenerife, Canary Islands, Spain.
Basilio
B
Valladares
University Institute of Tropical Diseases and Public Health of the Canary Islands, University of La Laguna, Avda. Astrofísico Fco. Sánchez, S/N, 38203 La Laguna, Tenerife, Canary Islands, Spain.
Jose E
JE
Piñero
University Institute of Tropical Diseases and Public Health of the Canary Islands, University of La Laguna, Avda. Astrofísico Fco. Sánchez, S/N, 38203 La Laguna, Tenerife, Canary Islands, Spain.
Jacob
J
Lorenzo-Morales
D048248:Acanthamoeba castellanii / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D002710:Chlorhexidine / Q000494:pharmacology; D004202:Disinfectants / Q000494:pharmacology; D005453:Fluorescence; D007202:Indicators and Reagents / Q000592:standards*; D020128:Inhibitory Concentration 50; D016014:Linear Models; D016015:Logistic Models; D010078:Oxazines / Q000592:standards*; D013997:Time Factors; D053058:Trophozoites / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D014966:Xanthenes / Q000592:standards*
D004202:Disinfectants; D007202:Indicators and Reagents; D010078:Oxazines; D014966:Xanthenes; C005843:resazurin; D002710:Chlorhexidine
Acanthamoeba; AlamarBlue®; In vitro assay; PrestoBlue®