ArticlePloS one2013
Multiplex qPCR for detection and absolute quantification of malaria.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 97 papers, 1 of them a synthesis that pooled it.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
97 citing papers in PubMed, 1 synthesis or guideline pooled it.
- Pooled it
- Trial
- Increase of Dose Associated With Decrease in Protection Against Controlled Human Malaria Infection by PfSPZ Vaccine in Tanzanian Adults.Clinical infectious diseases : an official publication of the Infectious Diseases Society of America · 2020Trial
- Trial
- Safety and Differential Antibody and T-Cell Responses to theThe American journal of tropical medicine and hygiene · 2019Trial
- Safety, Immunogenicity, and Protective Efficacy against Controlled Human Malaria Infection ofThe American journal of tropical medicine and hygiene · 2018Trial
- Trial
- Protection againstJCI insight · 2017Trial
- Measurement of parasitological data by quantitative real-time PCR from controlled human malaria infection trials at the Walter Reed Army Institute of Research.Malaria journal · 2014Trial
- Severe Plasmodium falciparum infection with multi-organ involvement in an asplenic patient: a case report.Malaria journal · 2026Article
- Phylogeographic dynamics of oropouche virus in the Colombian Amazon: Evolutionary insights in a climatic context.PLoS neglected tropical diseases · 2026Article
- Point of care performance of the improved BIOCREDIT Ag Pf/Pv (pLDH/pLDH) rapid diagnostic test: a cross-sectional study at a health center setting in Ethiopia.Malaria journal · 2026Article
- Accurate and sensitive TaqMan real-time PCR assays to detect and quantify simian malaria parasites Plasmodium cynomolgi, Plasmodium inui and Plasmodium coatneyi.Malaria journal · 2026Article
- Reversal of Chloroquine Resistance inInternational journal of molecular sciences · 2026Article
- The dynamics of asymptomatic Plasmodium spp. infections following 10 years of malaria control interventions in Northern Sahelian Ghana.PLoS neglected tropical diseases · 2026Observational
- Uncovering silent malaria reservoirs in Southern Ethiopia: community-based molecular detection of asymptomatic and sub-microscopic infections and their implications for diagnosis and control.Malaria journal · 2026Article
- High Burden of Febrile Sub-microscopic Plasmodium Mixed Infections in Central India: A Cross-Sectional Study.Infectious diseases and therapy · 2026Article
- Low prevalence ofFrontiers in epidemiology · 2026Article
- Article
- Global assessment of partial artemisinin resistance: multicenter trial across Kenya, Peru, and Thailand in patients with uncomplicated Plasmodium falciparum malaria.International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases · 2025Article
37 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We describe development of an absolute multiplex quantitative real-time PCR for detection of Plasmodium spp., P. falciparum and P. vivax targets in order to produce an assay amenable to high throughput but with reduced costs. Important qPCR experimental details and information that is critical to performance and reliability of assay results were investigated. Inhibition studies were performed to test and compare co-purification of PCR inhibitors in samples extracted from whole blood using either the manual or automated methods. To establish the most optimal qPCR reaction volume, volume titration of the reaction master mix was performed starting at 10 µl to 1 µl reaction master mix with 1 µl of template DNA in each reaction. As the reaction volume decreased, qPCR assays became more efficient with 1 µl reaction master mix being the most efficient. For more accurate quantification of parasites in a sample, we developed plasmid DNAs for all the three assay targets for absolute quantification. All of absolute qPCR assays performed with efficiency of more than 94%, R(2) values greater than 0.99 and the STDEV of each replicate was <0.167. Linear regression plots generated from absolute qPCR assays were used to estimate the corresponding parasite density from relative qPCR in terms of parasite/µl. One copy of plasmid DNA was established to be equivalent to 0.1 parasite/µl for Plasmodium spp. assay, 0.281 parasites for P. falciparum assay and 0.127 parasite/µl for P. vivax assay. This study demonstrates for the first time use of plasmid DNA in absolute quantification of malaria parasite. The use of plasmid DNA standard in quantification of malaria parasite will be critical as efforts are underway to harmonize molecular assays used in diagnosis of malaria.
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