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

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...
Ausführliche Beschreibung

Gespeichert in:

Autor*in:	Sykes, Melissa L [verfasserIn] Avery, Vicky M

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2009

Schlagwörter:	High Throughput Screening Pentamidine Human African Tropanosomiasis Final DMSO Concentration Alamar Blue Assay

Anmerkung:	© Sykes and Avery; licensee BioMed Central Ltd. 2009

Übergeordnetes Werk:	Enthalten in: Parasites & vectors - London : BioMed Central, 2008, 2(2009), 1 vom: 12. Nov.
Übergeordnetes Werk:	volume:2 ; year:2009 ; number:1 ; day:12 ; month:11

Links:	Volltext

DOI / URN:	10.1186/1756-3305-2-54

Katalog-ID:	SPR030165156

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520			\|a 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 $ IC_{50} $ 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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author_variant	m l s ml mls v m a vm vma
matchkey_str	article:17563305:2009----::lcfrsbsdibltasyoapeetoi34elomtohgtruhuwoeelcennotyaooa
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allfields	10.1186/1756-3305-2-54 doi (DE-627)SPR030165156 (SPR)1756-3305-2-54-e DE-627 ger DE-627 rakwb eng Sykes, Melissa L verfasserin aut 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 2009 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Sykes and Avery; licensee BioMed Central Ltd. 2009 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 $ IC_{50} $ 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. High Throughput Screening (dpeaa)DE-He213 Pentamidine (dpeaa)DE-He213 Human African Tropanosomiasis (dpeaa)DE-He213 Final DMSO Concentration (dpeaa)DE-He213 Alamar Blue Assay (dpeaa)DE-He213 Avery, Vicky M aut Enthalten in Parasites & vectors London : BioMed Central, 2008 2(2009), 1 vom: 12. Nov. (DE-627)558690076 (DE-600)2409480-8 1756-3305 nnns volume:2 year:2009 number:1 day:12 month:11 https://dx.doi.org/10.1186/1756-3305-2-54 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 2 2009 1 12 11
spelling	10.1186/1756-3305-2-54 doi (DE-627)SPR030165156 (SPR)1756-3305-2-54-e DE-627 ger DE-627 rakwb eng Sykes, Melissa L verfasserin aut 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 2009 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Sykes and Avery; licensee BioMed Central Ltd. 2009 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 $ IC_{50} $ 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. High Throughput Screening (dpeaa)DE-He213 Pentamidine (dpeaa)DE-He213 Human African Tropanosomiasis (dpeaa)DE-He213 Final DMSO Concentration (dpeaa)DE-He213 Alamar Blue Assay (dpeaa)DE-He213 Avery, Vicky M aut Enthalten in Parasites & vectors London : BioMed Central, 2008 2(2009), 1 vom: 12. Nov. (DE-627)558690076 (DE-600)2409480-8 1756-3305 nnns volume:2 year:2009 number:1 day:12 month:11 https://dx.doi.org/10.1186/1756-3305-2-54 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 2 2009 1 12 11
allfields_unstemmed	10.1186/1756-3305-2-54 doi (DE-627)SPR030165156 (SPR)1756-3305-2-54-e DE-627 ger DE-627 rakwb eng Sykes, Melissa L verfasserin aut 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 2009 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Sykes and Avery; licensee BioMed Central Ltd. 2009 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 $ IC_{50} $ 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. High Throughput Screening (dpeaa)DE-He213 Pentamidine (dpeaa)DE-He213 Human African Tropanosomiasis (dpeaa)DE-He213 Final DMSO Concentration (dpeaa)DE-He213 Alamar Blue Assay (dpeaa)DE-He213 Avery, Vicky M aut Enthalten in Parasites & vectors London : BioMed Central, 2008 2(2009), 1 vom: 12. Nov. (DE-627)558690076 (DE-600)2409480-8 1756-3305 nnns volume:2 year:2009 number:1 day:12 month:11 https://dx.doi.org/10.1186/1756-3305-2-54 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 2 2009 1 12 11
allfieldsGer	10.1186/1756-3305-2-54 doi (DE-627)SPR030165156 (SPR)1756-3305-2-54-e DE-627 ger DE-627 rakwb eng Sykes, Melissa L verfasserin aut 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 2009 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Sykes and Avery; licensee BioMed Central Ltd. 2009 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 $ IC_{50} $ 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. High Throughput Screening (dpeaa)DE-He213 Pentamidine (dpeaa)DE-He213 Human African Tropanosomiasis (dpeaa)DE-He213 Final DMSO Concentration (dpeaa)DE-He213 Alamar Blue Assay (dpeaa)DE-He213 Avery, Vicky M aut Enthalten in Parasites & vectors London : BioMed Central, 2008 2(2009), 1 vom: 12. Nov. (DE-627)558690076 (DE-600)2409480-8 1756-3305 nnns volume:2 year:2009 number:1 day:12 month:11 https://dx.doi.org/10.1186/1756-3305-2-54 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 2 2009 1 12 11
allfieldsSound	10.1186/1756-3305-2-54 doi (DE-627)SPR030165156 (SPR)1756-3305-2-54-e DE-627 ger DE-627 rakwb eng Sykes, Melissa L verfasserin aut 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 2009 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Sykes and Avery; licensee BioMed Central Ltd. 2009 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 $ IC_{50} $ 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. High Throughput Screening (dpeaa)DE-He213 Pentamidine (dpeaa)DE-He213 Human African Tropanosomiasis (dpeaa)DE-He213 Final DMSO Concentration (dpeaa)DE-He213 Alamar Blue Assay (dpeaa)DE-He213 Avery, Vicky M aut Enthalten in Parasites & vectors London : BioMed Central, 2008 2(2009), 1 vom: 12. Nov. (DE-627)558690076 (DE-600)2409480-8 1756-3305 nnns volume:2 year:2009 number:1 day:12 month:11 https://dx.doi.org/10.1186/1756-3305-2-54 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 2 2009 1 12 11
language	English
source	Enthalten in Parasites & vectors 2(2009), 1 vom: 12. Nov. volume:2 year:2009 number:1 day:12 month:11
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topic_facet	High Throughput Screening Pentamidine Human African Tropanosomiasis Final DMSO Concentration Alamar Blue Assay
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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 $ IC_{50} $ 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. 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author	Sykes, Melissa L
spellingShingle	Sykes, Melissa L misc High Throughput Screening misc Pentamidine misc Human African Tropanosomiasis misc Final DMSO Concentration misc Alamar Blue Assay 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
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topic_title	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 High Throughput Screening (dpeaa)DE-He213 Pentamidine (dpeaa)DE-He213 Human African Tropanosomiasis (dpeaa)DE-He213 Final DMSO Concentration (dpeaa)DE-He213 Alamar Blue Assay (dpeaa)DE-He213
topic	misc High Throughput Screening misc Pentamidine misc Human African Tropanosomiasis misc Final DMSO Concentration misc Alamar Blue Assay
topic_unstemmed	misc High Throughput Screening misc Pentamidine misc Human African Tropanosomiasis misc Final DMSO Concentration misc Alamar Blue Assay
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title	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
ctrlnum	(DE-627)SPR030165156 (SPR)1756-3305-2-54-e
title_full	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
author_sort	Sykes, Melissa L
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author_browse	Sykes, Melissa L Avery, Vicky M
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title_sort	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
title_auth	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
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 $ IC_{50} $ 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. © Sykes and Avery; licensee BioMed Central Ltd. 2009
abstractGer	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 $ IC_{50} $ 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. © Sykes and Avery; licensee BioMed Central Ltd. 2009
abstract_unstemmed	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 $ IC_{50} $ 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. © Sykes and Avery; licensee BioMed Central Ltd. 2009
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title_short	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
url	https://dx.doi.org/10.1186/1756-3305-2-54
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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

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