Development and optimization of a novel 384-well anti-malarial imaging assay validated for high-throughput screening.
Identifieur interne : 000816 ( Main/Exploration ); précédent : 000815; suivant : 000817Development 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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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Development and optimization of a novel 384-well anti-malarial imaging assay validated for high-throughput screening.</title><author><name sortKey="Duffy, Sandra" sort="Duffy, Sandra" uniqKey="Duffy S" first="Sandra" last="Duffy">Sandra Duffy</name><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></affiliation></author><author><name sortKey="Avery, Vicky M" sort="Avery, Vicky M" uniqKey="Avery V" first="Vicky M" last="Avery">Vicky M. Avery</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2012">2012</date><idno type="doi">10.4269/ajtmh.2012.11-0302</idno><idno type="RBID">pubmed:22232455</idno><idno type="pmid">22232455</idno><idno type="wicri:Area/PubMed/Corpus">000289</idno><idno type="wicri:Area/PubMed/Curation">000289</idno><idno type="wicri:Area/PubMed/Checkpoint">000272</idno><idno type="wicri:Area/Ncbi/Merge">000933</idno><idno type="wicri:Area/Ncbi/Curation">000933</idno><idno type="wicri:Area/Ncbi/Checkpoint">000933</idno><idno type="wicri:Area/Main/Merge">000812</idno><idno type="wicri:Area/Main/Curation">000816</idno><idno type="wicri:Area/Main/Exploration">000816</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Development and optimization of a novel 384-well anti-malarial imaging assay validated for high-throughput screening.</title><author><name sortKey="Duffy, Sandra" sort="Duffy, Sandra" uniqKey="Duffy S" first="Sandra" last="Duffy">Sandra Duffy</name><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></affiliation></author><author><name sortKey="Avery, Vicky M" sort="Avery, Vicky M" uniqKey="Avery V" first="Vicky M" last="Avery">Vicky M. Avery</name></author></analytic><series><title level="j">The American journal of tropical medicine and hygiene</title><idno type="e-ISSN">1476-1645</idno><imprint><date when="2012" type="published">2012</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Algorithms</term><term>Animals</term><term>Antimalarials (pharmacology)</term><term>Drug Discovery</term><term>Drug Resistance</term><term>Fluorescent Dyes (metabolism)</term><term>High-Throughput Screening Assays (methods)</term><term>Indoles (metabolism)</term><term>Microscopy, Confocal</term><term>Microscopy, Fluorescence</term><term>Parasitic Sensitivity Tests (methods)</term><term>Plasmodium falciparum (drug effects)</term><term>Plasmodium falciparum (growth & development)</term><term>Signal-To-Noise Ratio</term><term>Software</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Fluorescent Dyes</term><term>Indoles</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Antimalarials</term></keywords><keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Plasmodium falciparum</term></keywords><keywords scheme="MESH" qualifier="growth & development" xml:lang="en"><term>Plasmodium falciparum</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>High-Throughput Screening Assays</term><term>Parasitic Sensitivity Tests</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Algorithms</term><term>Animals</term><term>Drug Discovery</term><term>Drug Resistance</term><term>Microscopy, Confocal</term><term>Microscopy, Fluorescence</term><term>Signal-To-Noise Ratio</term><term>Software</term></keywords></textClass></profileDesc></teiHeader><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></front></TEI><affiliations><list><country><li>Australie</li></country></list><tree><noCountry><name sortKey="Avery, Vicky M" sort="Avery, Vicky M" uniqKey="Avery V" first="Vicky M" last="Avery">Vicky M. Avery</name></noCountry><country name="Australie"><noRegion><name sortKey="Duffy, Sandra" sort="Duffy, Sandra" uniqKey="Duffy S" first="Sandra" last="Duffy">Sandra Duffy</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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