A Plasmodium falciparum screening assay for anti-gametocyte drugs based on parasite lactate dehydrogenase detection

被引：96

作者：

D'Alessandro, Sarah ^{[1
]}

Silvestrini, Francesco ^{[2
]}

Dechering, Koen ^{[3
]}

Corbett, Yolanda ^{[1
]}

Parapini, Silvia ^{[1
]}

Timmerman, Martijn ^{[3
]}

Galastri, Laura ^{[4
]}

Basilico, Nicoletta ^{[5
]}

Sauerwein, Robert ^{[6
]}

Alano, Pietro ^{[2
]}

Taramelli, Donatella ^{[1
]}

机构：

[1] Univ Milan, Dipartimento Sci Farmacol & Biomol, I-20133 Milan, Italy

[2] Ist Super Sanita, Dipartimento Malattie Infett, I-00161 Rome, Italy

[3] TropIQ Hlth Sci, NL-6525 GA Nijmegen, Netherlands

[4] Assoc Italiana Volontari Sangue AVIS Comunale Mil, I-20133 Milan, Italy

[5] Univ Milan, Dipartimento Sci Biomed Chirurg & Odontoiatr, I-20133 Milan, Italy

[6] Radboud Univ Nijmegen, Dept Med Microbiol, Med Ctr, NL-6525 GA Nijmegen, Netherlands

来源：

JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY | 2013年 / 68卷 / 09期

基金：

比尔及梅琳达.盖茨基金会;

关键词：

malaria; gametocytes; drug screening; MALARIA; STAGE; TRANSMISSION; ARTESUNATE; PRIMAQUINE;

D O I：

10.1093/jac/dkt165

中图分类号：

R51 [传染病];

学科分类号：

100401 ;

摘要：

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. 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). 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. 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.

引用

页码：2048 / 2058

页数：11

共 35 条

[1] Quantitative assessment of Plasmodium falciparum sexual development reveals potent transmission-blocking activity by methylene blue [J].