Development and optimization of a novel 384-well anti-malarial imaging assay validated for high-throughput screening.
Identifieur interne : 000289 ( PubMed/Curation ); précédent : 000288; suivant : 000290Development 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
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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></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></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><pubmed><MedlineCitation Owner="NLM" Status="MEDLINE"><PMID Version="1">22232455</PMID><DateCreated><Year>2012</Year><Month>01</Month><Day>10</Day></DateCreated><DateCompleted><Year>2012</Year><Month>03</Month><Day>09</Day></DateCompleted><DateRevised><Year>2015</Year><Month>01</Month><Day>28</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Electronic">1476-1645</ISSN><JournalIssue CitedMedium="Internet"><Volume>86</Volume><Issue>1</Issue><PubDate><Year>2012</Year><Month>Jan</Month></PubDate></JournalIssue><Title>The American journal of tropical medicine and hygiene</Title><ISOAbbreviation>Am. J. Trop. Med. Hyg.</ISOAbbreviation></Journal><ArticleTitle>Development and optimization of a novel 384-well anti-malarial imaging assay validated for high-throughput screening.</ArticleTitle><Pagination><MedlinePgn>84-92</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.4269/ajtmh.2012.11-0302</ELocationID><Abstract><AbstractText>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.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Duffy</LastName><ForeName>Sandra</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Discovery Biology, Griffith University, Nathan, Australia. sandra.duffy@griffith.edu.au</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Avery</LastName><ForeName>Vicky M</ForeName><Initials>VM</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D023362">Evaluation Studies</PublicationType><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Am J Trop Med 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