Development and optimization of a novel 384-well anti-malarial imaging assay validated for high-throughput screening.
Identifieur interne : 000933 ( Ncbi/Checkpoint ); précédent : 000932; suivant : 000934Development and optimization of a novel 384-well anti-malarial imaging assay validated for high-throughput screening.
Auteurs : Sandra Duffy [Australie] ; Vicky M. AverySource :
- The American journal of tropical medicine and hygiene ; 2012.
English descriptors
- KwdEn :
- Algorithms, Animals, Antimalarials (pharmacology), Drug Discovery, Drug Resistance, Fluorescent Dyes (metabolism), High-Throughput Screening Assays (methods), Indoles (metabolism), Microscopy, Confocal, Microscopy, Fluorescence, Parasitic Sensitivity Tests (methods), Plasmodium falciparum (drug effects), Plasmodium falciparum (growth & development), Signal-To-Noise Ratio, Software.
- MESH :
- chemical , metabolism : Fluorescent Dyes, Indoles.
- chemical , pharmacology : Antimalarials.
- drug effects : Plasmodium falciparum.
- growth & development : Plasmodium falciparum.
- methods : High-Throughput Screening Assays, Parasitic Sensitivity Tests.
- Algorithms, Animals, Drug Discovery, Drug Resistance, Microscopy, Confocal, Microscopy, Fluorescence, Signal-To-Noise Ratio, Software.
Abstract
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.
DOI: 10.4269/ajtmh.2012.11-0302
PubMed: 22232455
Affiliations:
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<affiliation wicri:level="1"><nlm:affiliation>Discovery Biology, Griffith University, Nathan, Australia. sandra.duffy@griffith.edu.au</nlm:affiliation>
<country xml:lang="fr">Australie</country>
<wicri:regionArea>Discovery Biology, Griffith University, Nathan</wicri:regionArea>
<wicri:noRegion>Nathan</wicri:noRegion>
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<author><name sortKey="Avery, Vicky M" sort="Avery, Vicky M" uniqKey="Avery V" first="Vicky M" last="Avery">Vicky M. Avery</name>
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<term>Drug Resistance</term>
<term>Fluorescent Dyes (metabolism)</term>
<term>High-Throughput Screening Assays (methods)</term>
<term>Indoles (metabolism)</term>
<term>Microscopy, Confocal</term>
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<front><div type="abstract" xml:lang="en">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.</div>
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