A Plasmodium falciparum screening assay for anti-gametocyte drugs based on parasite lactate dehydrogenase detection
Articolo
Data di Pubblicazione:
2013
Citazione:
A Plasmodium falciparum screening assay for anti-gametocyte drugs based on parasite lactate dehydrogenase detection / S. D’Alessandro, F. Silvestrini, K. Dechering, Y. Corbett, S. Parapini, M. Timmerman, L. Galastri, N. Basilico, R. Sauerwein, P. Alano, D. Taramelli. - In: JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY. - ISSN 0305-7453. - 68:9(2013 Sep), pp. 2048-2058. [10.1093/jac/dkt165]
Abstract:
Objectives: Plasmodium gametocytes, responsible for malaria parasite transmission from humans to mosquitoes,
represent a crucial target for new antimalarial drugs to achieve malaria elimination/eradication. We developed a
novel colorimetric screening method for anti-gametocyte compounds based on the parasite lactate dehydrogenase
(pLDH) assay, already standardized for asexual stages, to measure gametocyte viability and drug susceptibility.
Methods: Gametocytogenesis of 3D7 and NF54 Plasmodium falciparum strains was induced in vitro and asexual
parasites were depleted with N-acetylglucosamine. Gametocytes were treated with dihydroartemisinin, epoxomicin, methylene blue, primaquine, puromycin or chloroquine in 96-well plates and the pLDH activity was evaluated using a modified Makler protocol. Mosquito infectivity was measured by the standard membrane feeding assay (SMFA).
Results: A linear correlation was found between gametocytaemia determined by Giemsa staining and pLDH activity.
A concentration-dependent reduction in pLDH activity was observed after 72 h of drug treatment, whereas an
additional 72 h of incubation without drugs was required to obtain complete inhibition of gametocyte viability.
SMFA on treated and control gametocytes confirmed that a reduction in pLDH activity translates into reduced
oocyst development in the mosquito vector.
Conclusions: The gametocyte pLDH assay is fast, easy to perform, cheap and reproducible and is suitable for
screening novel transmission-blocking compounds, which does not require parasite transgenic lines.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
malaria; gametocytes; drug screening
Elenco autori:
S. D’Alessandro, F. Silvestrini, K. Dechering, Y. Corbett, S. Parapini, M. Timmerman, L. Galastri, N. Basilico, R. Sauerwein, P. Alano, D. Taramelli
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