ArticleMalaria journal2012
A high-throughput method to detect Plasmodium falciparum clones in limiting dilution microplates.
Article in Malaria journal, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed, 24 citations in OpenAlex.
- Article
- Development of a single-cell cloning technique for isolation of Pentatrichomonas hominis: a promising tool for diagnosing Trichomonas spp. infections in the pig breeding industry.Parasites & vectors · 2025Article
- M-iScience · 2024Article
- Cultivation of Asexual Intraerythrocytic Stages ofPathogens (Basel, Switzerland) · 2023Review
- Conditional permeabilization of the P. falciparum plasma membrane in infected cells links cation influx to reduced membrane integrity.PloS one · 2023Article
- Roles of the RON3 C-terminal fragment in erythrocyte invasion and blood-stage parasite proliferation inFrontiers in cellular and infection microbiology · 2023Article
- Chemical Biology Tools To Investigate Malaria Parasites.Chembiochem : a European journal of chemical biology · 2021Review
- Improved Plasmodium falciparum dilution cloning through efficient quantification of parasite numbers and c-SNARF detection.Malaria journal · 2021Article
- Complex nutrient channel phenotypes despite Mendelian inheritance in a Plasmodium falciparum genetic cross.PLoS pathogens · 2020Article
- Article
- Article
- A CLAG3 mutation in an amphipathic transmembrane domain alters malaria parasite nutrient channels and confers leupeptin resistance.Infection and immunity · 2015Article
- Why do malaria parasites increase host erythrocyte permeability?Trends in parasitology · 2014Review
- Proteolysis at a specific extracellular residue implicates integral membrane CLAG3 in malaria parasite nutrient channels.PloS one · 2014Article
- An epigenetic antimalarial resistance mechanism involving parasite genes linked to nutrient uptake.The Journal of biological chemistry · 2013Article
- Malaria parasites tolerate a broad range of ionic environments and do not require host cation remodelling.Molecular microbiology · 2013Article
- Insights gained from P. falciparum cultivation in modified media.TheScientificWorldJournal · 2013Review
- Solute restriction reveals an essential role for clag3-associated channels in malaria parasite nutrient acquisition.Molecular pharmacology · 2012Article
Corrections and comments
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Authors and funding
5 authors at 3 institutions in 2 countries.
Funding
Abstract
backgroundMolecular and cellular studies of Plasmodium falciparum require cloning of parasites by limiting dilution cultivation, typically performed in microplates. The parasite's slow replication rate combined with laborious methods for identification of positive wells has limited these studies. A new high-throughput method for detecting growth without compromising parasite viability is reported.
methodsIn vitro parasite cultivation is associated with extracellular acidification. A survey of fluorescent pH indicators identified 5-(and-6)-carboxy SNARF-1 as a membrane-impermeant dye with a suitable pKa value. Conditions for facile detection of viable parasites in 96-well microplates were optimized and used for limiting dilution cloning of genetic cross progeny and transfected parasites.
results5-(and-6)-carboxy SNARF-1 is a two-emission wavelength dye that accurately reported extracellular pH in parasite cultures. It readily detected parasite growth in microplate wells and yielded results comparable to labour-intensive examination of Giemsa-stained smears. The dye is non-toxic, allowing parasite detection without transfer of culture material to additional plates for separate assays. This dye was used with high-throughput limiting dilution culture to generate additional progeny clones from the HB3 × Dd2 genetic cross.
conclusionsThis fluorescence-based assay represents a low-cost, efficient method for detection of viable parasites in microplate wells; it can be easily expanded by automation.
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