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基于 SYBR Green I、PicoGreen 和 [3H]-次黄嘌呤的体外抗疟筛选方法比较尼日利亚传统医学植物。

Comparison of SYBR Green I-, PicoGreen-, and [3H]-hypoxanthine-based assays for in vitro antimalarial screening of plants from Nigerian ethnomedicine.

机构信息

Department of Pharmacology and Therapeutics, College of Health Sciences, Ladoke Akintola University of Technology, Oshogbo, Nigeria.

出版信息

Parasitol Res. 2010 Mar;106(4):933-9. doi: 10.1007/s00436-010-1743-z. Epub 2010 Feb 18.

Abstract

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 green (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 [(3)H]-hypoxanthine (HP) incorporation assays for in vitro antimalarial screening of medicinal plants. The 50% inhibitory concentration (IC(50)) 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 micro-flourimetric 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.

摘要

体外抗疟药物筛选的标准方法基于同位素测定法,该方法昂贵,并且使用放射性材料,在发展中国家存在有限的可用性、安全性和处理问题。已经报道了使用非放射性 DNA 染料 SYBR Green I (SG) 和 PICO green (PG) 进行抗疟筛选。然而,尚未比较这两种 DNA 染料在药用植物抗疟筛选中的应用。因此,本研究比较了 SG、PG 与 [(3)H]-次黄嘌呤 (HP) 掺入测定法,用于药用植物的体外抗疟筛选。三种方法获得的药用植物和标准抗疟药物抗疟活性的 50%抑制浓度 (IC(50)) 值相似。本研究产生的数据表明,非放射性微量荧光测定法足够灵敏,可从缺乏活性的植物提取物中重现性地识别具有抗疟活性的植物提取物。与 SG 的信噪比为 7 和 PG 的信噪比为 8 相比,基于 HP 的测定法的信号与噪声比最高为 100。基于 SG 的测定法比基于 PG 和 HP 的测定法更便宜。SG 似乎是抗疟药物筛选的一种具有成本效益的替代方法,也是一种可行的技术,特别是在发展中国家,可能有助于抗疟药物发现过程。

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