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 Plasmodium falciparum 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-ɛ-(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.
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机译：放射性同位素测定涉及费用，多步骤方案，设备和放射性安全性要求，这些要求在高通量药物测试中存在问题。这项研究报告了一种可用于抗疟疾药物筛选的替代方法，简单，可靠，便宜的一步荧光检测方法。寄生虫的生长通过使用SYBR Green I（一种与疟原虫DNA接触后荧光增强明显的染料）来确定。通过测试已知的抗恶性疟原虫菌株D6的抗疟药，进行了这种荧光测定法与标准放射性同位素方法的并排比较。两种测定方法均用于确定药物的有效浓度，该浓度可在每种药物存在的情况下，在寄生虫生长48小时后将观察到的计数（EC50）降低50％。两种技术的氯喹，奎宁，甲氟喹，青蒿素和3,6-双-（-N，N-二乙氨基）-戊氧基黄酮的EC50相似或相同。用这种新的荧光测定法获得的结果表明，它可能是高通量抗疟药物筛选的理想方法。
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