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Sykes ML, Avery VM. A luciferase based viability assay for ATP detection in 384-well format for high throughput whole cell screening of Trypanosoma brucei brucei bloodstream form strain 427. Parasit Vectors. 2009;2(1):54doi: 10.1186/1756-3305-2-54.
Sykes, M. L., & Avery, V. M. (2009). A luciferase based viability assay for ATP detection in 384-well format for high throughput whole cell screening of Trypanosoma brucei brucei bloodstream form strain 427. Parasites & vectors, 2(1), 54. https://doi.org/10.1186/1756-3305-2-54
Sykes, Melissa L, and Avery, Vicky M. "A luciferase based viability assay for ATP detection in 384-well format for high throughput whole cell screening of Trypanosoma brucei brucei bloodstream form strain 427." Parasites & vectors vol. 2,1 (2009): 54. doi: https://doi.org/10.1186/1756-3305-2-54
Sykes ML, Avery VM. A luciferase based viability assay for ATP detection in 384-well format for high throughput whole cell screening of Trypanosoma brucei brucei bloodstream form strain 427. Parasit Vectors. 2009 Nov 12;2(1):54. doi: 10.1186/1756-3305-2-54. PMID: 19909542; PMCID: PMC2781010.
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Parasit Vectors. 2009 Nov 12;2(1):54. doi: 10.1186/1756-3305-2-54.

A luciferase based viability assay for ATP detection in 384-well format for high throughput whole cell screening of Trypanosoma brucei brucei bloodstream form strain 427.

Parasites & vectors

Melissa L Sykes, Vicky M Avery

Affiliations

  1. Eskitis Institute for Cell and Molecular Therapies, Griffith University, Eskitis Building N27, Brisbane Innovation Park, Don Young Road, Nathan, Queensland, Australia. [email protected].

PMID: 19909542 PMCID: PMC2781010 DOI: 10.1186/1756-3305-2-54

Abstract

BACKGROUND: Human African Trypanosomiasis (HAT) is caused by two trypanosome species, Trypanosoma brucei rhodesiense and Trypanosoma brucei gambiense. Current drugs available for the treatment of HAT have significant issues related to toxicity, administration regimes with limited effectiveness across species and disease stages, thus there is a considerable need to find alternative drugs. A well recognised approach to identify new drug candidates is high throughput screening (HTS) of large compound library collections.

RESULTS: We describe here the development of a luciferase based viability assay in 384-well plate format suitable for HTS of T.b.brucei. The parameters that were explored to determine the final HTS assay conditions are described in detail and include DMSO tolerability, Z', diluents and cell inoculum density. Reference compound activities were determined for diminazene, staurosporine and pentamidine and compared to previously published IC50 data obtained. The assay has a comparable sensitivity to reference drugs and is more cost effective than the 96-well format currently reported for T.b.brucei.

CONCLUSION: Due to the reproducibility and sensitivity of this assay it is recommended for potential HTS application. As it is commercially available this assay can also be utilised in many laboratories for both large and small scale screening.

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