Comparison of SYBR Green I-, PicoGreen-, and [3H]-hypoxanthine-based assays for in vitro antimalarial screening of plants from Nigerian ethnomedicine
Comparison of SYBR Green I-, PicoGreen-, and [3H]-hypoxanthine-based assays for in vitro antimalarial screening of plants from Nigerian ethnomedicine
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DOI：
10.1007/s00436-010-1743-z
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发表时间：
2010-03-01
影响因子：
2
通讯作者：
Oduola, Ayoade M. J.
中科院分区：
文献类型：
--
作者：
Abiodun, Oyindamola O.;Gbotosho, Grace O.;Oduola, Ayoade M. J.
体外抗疟疾药物筛选的标准方法是基于同位素分析，这种方法价格昂贵，使用的放射性材料有限，在发展中国家存在可获得性、安全性和处置问题。已有报道使用非放射性DNA染色SYBR Green I(SG)和Pico Greena(R)(PG)进行抗疟筛查。然而，这两种DNA染色在药用植物抗疟筛查中的应用还没有进行过比较。因此，这项研究比较了SG、PG和[H-3]-次黄嘌呤(HP)掺入试验在药用植物体外抗疟疾筛选中的作用。用三种方法测得的药用植物和标准抗疟药物的半数抑制浓度(IC50)值相近。这项研究产生的数据表明，非放射性微量荧光法具有足够的灵敏度，可以重复地识别具有抗疟疾活性的植物提取物与缺乏活性的植物提取物。基于HP的检测显示出最稳健的信噪比为100，而SG和PG的信噪比分别为7和8。基于SG的检测比基于PG和HP的检测更便宜。SG似乎是抗疟疾药物筛选的一种具有成本效益的替代办法，也是一种可行的技术，可以促进抗疟疾药物的发现过程，特别是在发展中国家。
The standard method for in vitro antimalarial drug screening is based on the isotopic assay which is expensive and utilizes radioactive materials with limited availability, safety, and disposal problems in developing countries. The use of non-radioactive DNA stains SYBR Green I (SG) and PICO greenA (R) (PG) for antimalarial screening had been reported. However, the use of the two DNA stains for antimalarial screening of medicinal plants has not been compared. Thus, this study compared SG, PG with the [H-3]-hypoxanthine (HP) incorporation assays for in vitro antimalarial screening of medicinal plants. The 50% inhibitory concentration (IC50) values obtained using the three methods for antimalarial activity of medicinal plants and standard antimalarial drugs were similar. Data generated from this study suggests that the non-radioactive microflourimetric assay is sufficiently sensitive to reproducibly identify plant extracts with antimalarial activity from those lacking activity. The HP-based assay exhibited the most robust signal-to-noise ratio of 100, compared with signal-to-noise ratios of 7 for SG and 8 for PG. The SG-based assay is less expensive than the PG- and HP-based assays. SG appears to be a cost-effective alternative for antimalarial drug screening and a viable technique that may facilitate antimalarial drug discovery process especially in developing countries.