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 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.
Sandra
S
Duffy
Vicky M
VM
Avery
D000465:Algorithms; D000818:Animals; D000962:Antimalarials / Q000494:pharmacology*; D055808:Drug Discovery; D004351:Drug Resistance; D005456:Fluorescent Dyes / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D007211:Indoles / Q000378:metabolism; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D021261:Parasitic Sensitivity Tests / Q000379:methods*; D010963:Plasmodium falciparum / Q000187:drug effects* / Q000254:growth & development*; D059629:Signal-To-Noise Ratio; D012984:Software
D000962:Antimalarials; D005456:Fluorescent Dyes; D007211:Indoles; C007293:DAPI