ArticleMalaria journal2011
An improved method for undertaking limiting dilution assays for in vitro cloning of Plasmodium falciparum parasites.
Article in Malaria journal, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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10 citing papers in PubMed, 16 citations in OpenAlex.
- 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
- Expanding the fluorescent toolkit: Blue fluorescent protein-expressing Plasmodium berghei for enhanced multiplex microscopy.PloS one · 2025Article
- Cultivation of Asexual Intraerythrocytic Stages ofPathogens (Basel, Switzerland) · 2023Review
- Uncovering the role of Rad51 in homologous recombination-mediated antigenic diversification in the human malaria parasiteFrontiers in molecular biosciences · 2023Article
- A bioreactor system for the manufacture of a genetically modified Plasmodium falciparum blood stage malaria cell bank for use in a clinical trial.Malaria journal · 2018Article
- Article
- Highly Sensitive Two-Dimensional Paper Network Incorporating Biotin-Streptavidin for the Detection of Malaria.Analytical chemistry · 2016Article
- Highly sensitive immunoassay based on controlled rehydration of patterned reagents in a 2-dimensional paper network.Analytical chemistry · 2014Article
- A high-throughput method to detect Plasmodium falciparum clones in limiting dilution microplates.Malaria journal · 2012Article
- Modelling the dynamics of Plasmodium falciparum histidine-rich protein 2 in human malaria to better understand malaria rapid diagnostic test performance.Malaria journal · 2012Article
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundObtaining single parasite clones is required for many techniques in malaria research. Cloning by limiting dilution using microscopy-based assessment for parasite growth is an arduous and labor-intensive process. An alternative method for the detection of parasite growth in limiting dilution assays is using a commercial ELISA histidine-rich protein II (HRP2) detection kit.
methodsDetection of parasite growth was undertaken using HRP2 ELISA and compared to thick film microscopy. An HRP2 protein standard was used to determine the detection threshold of the HRP2 ELISA assay, and a HRP2 release model was used to extrapolate the amount of parasite growth required for a positive result.
resultsThe HRP2 ELISA was more sensitive than microscopy for detecting parasite growth. The minimum level of HRP2 protein detection of the ELISA was 0.11 ng/ml. Modeling of HRP2 release determined that 2,116 parasites are required to complete a full erythrocytic cycle to produce sufficient HRP2 to be detected by the ELISA. Under standard culture conditions this number of parasites is likely to be reached between 8 to 14 days of culture.
conclusionsThis method provides an accurate and simple way for the detection of parasite growth in limiting dilution assays, reducing time and resources required in traditional methods. Furthermore the method uses spent culture media instead of the parasite-infected red blood cells, enabling culture to continue.
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