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量子点:抗疟药物检测的新工具。

Quantum dots: a new tool for anti-malarial drug assays.

机构信息

Center for Neglected Diseases Drug Discovery, Institut Pasteur Korea, Seongnam-si, Gyeonggi-do, South Korea.

出版信息

Malar J. 2011 May 9;10:118. doi: 10.1186/1475-2875-10-118.

Abstract

BACKGROUND

Malaria infects over 300 million people every year and one of the major obstacles for the eradication of the disease is parasite's resistance to current chemotherapy, thus new drugs are urgently needed. Quantum dot (QD) is a fluorescent nanocrystal that has been in the spotlight as a robust tool for visualization of live cell processes in real time. Here, a simple and efficient method using QD to directly label Plasmodium falciparum-infected erythrocytes (iRBCs) was searched in order to use the QD as a probe in an anti-malarial drug-screening assay.

METHODS

A range of QDs with different chemical coatings were tested for their ability to specifically bind iRBCs by immunofluorescence assay (IFA). One QD was selected and used to detect parasite growth and drug sensitivity by flow cytometry.

RESULTS

PEGylated-cationic QD (PCQD) was found to specifically label infected erythrocytes preferentially with late stage parasites. The detection of QD-labelled infected erythrocytes by flow cytometry was sensitive enough to monitor chloroquine anti-malarial toxicity with a drug incubation period as short as 24 h (EC50 = 113nM). A comparison of our assay with another widely used anti-malarial drug screening assay, the pLDH assay, showed that PCQD-based assay had 50% improved sensitivity in detecting drug efficacy within a parasite life cycle. An excellent Z-factor of 0.8 shows that the QD assay is suitable for high-throughput screening.

CONCLUSIONS

This new assay can offer a rapid and robust platform to screen novel classes of anti-malarial drugs.

摘要

背景

每年有超过 3 亿人感染疟疾,而该疾病根除的主要障碍之一是寄生虫对当前化疗药物的耐药性,因此急需新的药物。量子点(QD)是一种荧光纳米晶体,已成为实时可视化活细胞过程的强大工具。在这里,我们搜索了一种使用 QD 直接标记恶性疟原虫感染红细胞(iRBC)的简单有效的方法,以便将 QD 用作抗疟药物筛选测定中的探针。

方法

通过免疫荧光法(IFA)测试了一系列具有不同化学涂层的 QD,以评估其与 iRBC 特异性结合的能力。选择了一种 QD 并用于通过流式细胞术检测寄生虫的生长和药物敏感性。

结果

发现聚乙二醇化阳离子 QD(PCQD)特异性标记感染的红细胞,优先标记晚期寄生虫。通过流式细胞术检测 QD 标记的感染红细胞的灵敏度足以监测氯喹抗疟毒性,药物孵育时间短至 24 小时(EC50=113nM)。与另一种广泛使用的抗疟药物筛选测定法 pLDH 测定法的比较表明,基于 PCQD 的测定法在检测寄生虫生命周期内药物疗效方面的灵敏度提高了 50%。出色的 Z 因子为 0.8 表明 QD 测定法适合高通量筛选。

结论

这种新的测定法可以提供一种快速而强大的平台,用于筛选新型抗疟药物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/863f/3112454/1e43b5fbec30/1475-2875-10-118-1.jpg

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