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

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

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

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

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

Authors

Annemarie M Voorberg-van der Wel¹, Anne-Marie Zeeman¹, Ivonne G Nieuwenhuis¹, Nicole M van der Werff¹, Clemens H M Kocken²

Affiliations

¹ Department of Parasitology, Biomedical Primate Research Centre, Rijswijk, The Netherlands.
² Department of Parasitology, Biomedical Primate Research Centre, Rijswijk, The Netherlands. kocken@bprc.nl.

PMID: 35821489
DOI: 10.1007/978-1-0716-2453-1_31

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.

Keywords: Bioluminescence (BL); Drug screening; Dual; Firefly luciferase; Hepatocytes; Hypnozoites; Liver; Liver stages; Malaria; Monkeys; NanoLuc; Plasmodium cynomolgi; Sporozoites.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Hepatocytes
Humans
Luciferases / genetics
Malaria* / diagnosis
Malaria* / parasitology
Plasmodium cynomolgi* / physiology
Recurrence

Substances

Luciferases
nanoluc