TY - JOUR
T1 - Simple and Inexpensive Fluorescence-Based Technique for High-Throughput Antimalarial Drug Screening
AU - Smilkstein, Martin
AU - Sriwilaijaroen, Nongluk
AU - Kelly, Jane Xu
AU - Wilairat, Prapon
AU - Riscoe, Michael
PY - 2004/5
Y1 - 2004/5
N2 - Radioisotopic assays involve expense, multistep protocols, equipment, and radioactivity safety requirements which are problematic in high-throughput drug testing. This study reports an alternative, simple, robust, inexpensive, one-step fluorescence assay for use in antimalarial drug screening. Parasite growth is determined by using SYBR Green I, a dye with marked fluorescence enhancement upon contact with Plasmodium DNA. A side-by-side comparison of this fluorescence assay and a standard radioisotopic method was performed by testing known antimalarial agents against Plasmodiumfalciparum strain D6. Both assay methods were used to determine the effective concentration of drug that resulted in a 50% reduction in the observed counts (EC50) after 48 h of parasite growth in the presence of each drug. The EC50S of chloroquine, quinine, mefloquine, artemisinin, and 3,6-bis-ε -(N,N-diethylamino)-amyloxyxanthone were similar or identical by both techniques. The results obtained with this new fluorescence assay suggest that it may be an ideal method for high-throughput antimalarial drug screening.
AB - Radioisotopic assays involve expense, multistep protocols, equipment, and radioactivity safety requirements which are problematic in high-throughput drug testing. This study reports an alternative, simple, robust, inexpensive, one-step fluorescence assay for use in antimalarial drug screening. Parasite growth is determined by using SYBR Green I, a dye with marked fluorescence enhancement upon contact with Plasmodium DNA. A side-by-side comparison of this fluorescence assay and a standard radioisotopic method was performed by testing known antimalarial agents against Plasmodiumfalciparum strain D6. Both assay methods were used to determine the effective concentration of drug that resulted in a 50% reduction in the observed counts (EC50) after 48 h of parasite growth in the presence of each drug. The EC50S of chloroquine, quinine, mefloquine, artemisinin, and 3,6-bis-ε -(N,N-diethylamino)-amyloxyxanthone were similar or identical by both techniques. The results obtained with this new fluorescence assay suggest that it may be an ideal method for high-throughput antimalarial drug screening.
UR - http://www.scopus.com/inward/record.url?scp=2142640849&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=2142640849&partnerID=8YFLogxK
U2 - 10.1128/AAC.48.5.1803-1806.2004
DO - 10.1128/AAC.48.5.1803-1806.2004
M3 - Article
C2 - 15105138
AN - SCOPUS:2142640849
SN - 0066-4804
VL - 48
SP - 1803
EP - 1806
JO - Antimicrobial agents and chemotherapy
JF - Antimicrobial agents and chemotherapy
IS - 5
ER -