基于双荧光素酶的疟疾潜隐子快速灵敏检测用于抗复发化合物的发现

Dual-Luciferase-Based Fast and Sensitive Detection of Malaria Hypnozoites for the Discovery of Anti-relapse Compounds.

作者信息

Voorberg-van der Wel Annemarie M, Zeeman Anne-Marie, Nieuwenhuis Ivonne G, van der Werff Nicole M, Kocken Clemens H M

机构信息

Department of Parasitology, Biomedical Primate Research Centre, Rijswijk, The Netherlands.

出版信息

Methods Mol Biol. 2022;2524:397-408. doi: 10.1007/978-1-0716-2453-1_31.

DOI:10.1007/978-1-0716-2453-1_31

PMID:35821489

Abstract

Malaria hypnozoites are dormant parasite stages that reside inside hepatocytes. Upon activation, these stages can resume growth, causing new episodes of blood stage malaria infection. This chapter describes a fast and sensitive protocol for the detection of bioluminescent (BL) hypnozoites in vitro. Using transgenic Plasmodium cynomolgi parasites that differentially express the BL reporter proteins firefly luciferase and the ultrabright NanoLuc, hypnozoites can be distinguished from liver stage schizonts. This robust method sets the stage for implementation in large-scale drug screening platforms with the aim to find new compounds that eliminate hypnozoites.

摘要

疟原虫休眠子是存在于肝细胞内的休眠寄生虫阶段。激活后，这些阶段可恢复生长，导致新的血液阶段疟疾感染发作。本章描述了一种在体外检测生物发光（BL）休眠子的快速灵敏方法。利用差异表达BL报告蛋白萤火虫荧光素酶和超亮纳米荧光素的转基因食蟹猴疟原虫寄生虫，休眠子可与肝期裂殖体区分开来。这种可靠的方法为在大规模药物筛选平台上实施奠定了基础，旨在寻找消除休眠子的新化合物。