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Development and Optimization of a Novel 384-Well Anti-Malarial Imaging Assay Validated for High-Throughput Screening

With the increasing occurrence of drug resistance in the malaria parasite, Plasmodium falciparum, there is a great need for new and novel anti-malarial drugs. We have developed a 384-well, high-throughput imaging assay for the detection of new anti-malarial compounds, which was initially validated b...

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Detalles Bibliográficos
Autores principales: Duffy, Sandra, Avery, Vicky M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society of Tropical Medicine and Hygiene 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3247113/
https://www.ncbi.nlm.nih.gov/pubmed/22232455
http://dx.doi.org/10.4269/ajtmh.2012.11-0302
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author Duffy, Sandra
Avery, Vicky M.
author_facet Duffy, Sandra
Avery, Vicky M.
author_sort Duffy, Sandra
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description With the increasing occurrence of drug resistance in the malaria parasite, Plasmodium falciparum, there is a great need for new and novel anti-malarial drugs. We have developed a 384-well, high-throughput imaging assay for the detection of new anti-malarial compounds, which was initially validated by screening a marine natural product library, and subsequently used to screen more than 3 million data points from a variety of compound sources. Founded on another fluorescence-based P. falciparum growth inhibition assay, the DNA-intercalating dye 4′,6-diamidino-2-phenylindole, was used to monitor changes in parasite number. Fluorescent images were acquired on the PerkinElmer Opera High Throughput confocal imaging system and analyzed with a spot detection algorithm using the Acapella data processing software. Further optimization of this assay sought to increase throughput, assay stability, and compatibility with our high-throughput screening equipment platforms. The assay typically yielded Z'-factor values of 0.5–0.6, with signal-to-noise ratios of 12.
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spelling pubmed-32471132012-01-10 Development and Optimization of a Novel 384-Well Anti-Malarial Imaging Assay Validated for High-Throughput Screening Duffy, Sandra Avery, Vicky M. Am J Trop Med Hyg Articles With the increasing occurrence of drug resistance in the malaria parasite, Plasmodium falciparum, there is a great need for new and novel anti-malarial drugs. We have developed a 384-well, high-throughput imaging assay for the detection of new anti-malarial compounds, which was initially validated by screening a marine natural product library, and subsequently used to screen more than 3 million data points from a variety of compound sources. Founded on another fluorescence-based P. falciparum growth inhibition assay, the DNA-intercalating dye 4′,6-diamidino-2-phenylindole, was used to monitor changes in parasite number. Fluorescent images were acquired on the PerkinElmer Opera High Throughput confocal imaging system and analyzed with a spot detection algorithm using the Acapella data processing software. Further optimization of this assay sought to increase throughput, assay stability, and compatibility with our high-throughput screening equipment platforms. The assay typically yielded Z'-factor values of 0.5–0.6, with signal-to-noise ratios of 12. The American Society of Tropical Medicine and Hygiene 2012-01-01 /pmc/articles/PMC3247113/ /pubmed/22232455 http://dx.doi.org/10.4269/ajtmh.2012.11-0302 Text en ©The American Society of Tropical Medicine and Hygiene Freely available online through the American Journal of Tropical Medicine and Hygiene Open Access option.
spellingShingle Articles
Duffy, Sandra
Avery, Vicky M.
Development and Optimization of a Novel 384-Well Anti-Malarial Imaging Assay Validated for High-Throughput Screening
title Development and Optimization of a Novel 384-Well Anti-Malarial Imaging Assay Validated for High-Throughput Screening
title_full Development and Optimization of a Novel 384-Well Anti-Malarial Imaging Assay Validated for High-Throughput Screening
title_fullStr Development and Optimization of a Novel 384-Well Anti-Malarial Imaging Assay Validated for High-Throughput Screening
title_full_unstemmed Development and Optimization of a Novel 384-Well Anti-Malarial Imaging Assay Validated for High-Throughput Screening
title_short Development and Optimization of a Novel 384-Well Anti-Malarial Imaging Assay Validated for High-Throughput Screening
title_sort development and optimization of a novel 384-well anti-malarial imaging assay validated for high-throughput screening
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3247113/
https://www.ncbi.nlm.nih.gov/pubmed/22232455
http://dx.doi.org/10.4269/ajtmh.2012.11-0302
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