| Author | Vera-Arias, Claudia A. | |
| Author | Holzschuh, Aurel | |
| Author | Oduma, Colins O | |
| Author | Badu, Kingsley | |
| Author | Abdul-Hakim, Mutala | |
| Author | Yukich, Joshua | |
| Author | Hetzel, Manuel W | |
| Author | Fakih, Bakar S | |
| Author | Ali, Abdullah | |
| Author | Ferreira, Marcelo U | |
| Author | Ladeia-Andrade, Simone | |
| Author | Sáenz, Fabián E | |
| Author | Afrane, Yaw | |
| Author | Zemene, Endalew | |
| Author | Yewhalaw, Delenasaw | |
| Author | Kazura, James W | |
| Author | Yan, Guiyun | |
| Author | Koepfli, Cristian | |
| Access date | 2022-12-18T12:05:41Z | |
| Available date | 2022-12-18T12:05:41Z | |
| Document date | 2022 | |
| Citation | VERA-ARIAS, Claudia A. et al. High-throughput Plasmodium falciparum hrp2 and hrp3 gene deletion typing by digital PCR to monitor malaria rapid diagnostic test efficacy. eLife, v.11, e72083, p. 1 - 16, June 2022. | en_US |
| ISSN | 2050-084X | en_US |
| URI | https://www.arca.fiocruz.br/handle/icict/56029 | |
| Language | eng | en_US |
| Publisher | Bavesh D Kana, University of the Witwatersrand, South Africa | en_US |
| Rights | open access | |
| Subject in Portuguese | Plasmodium falciparum | en_US |
| Subject in Portuguese | Alto rendimento | en_US |
| Subject in Portuguese | Tipagem de deleção dos genes hrp2 e hrp3 | en_US |
| Subject in Portuguese | PCR digital | en_US |
| Subject in Portuguese | Eficácia do teste de diagnóstico | en_US |
| Subject in Portuguese | Para monitorar a malária rapidamente | en_US |
| Title | High-throughput Plasmodium falciparum hrp2 and hrp3 gene deletion typing by digital PCR to monitor malaria rapid diagnostic test efficacy | en_US |
| Type | Article | |
| DOI | 10.7554/eLife.72083 | |
| Abstract | Most rapid diagnostic tests for Plasmodium falciparum malaria target the Histidine-Rich
Proteins 2 and 3 (HRP2 and HRP3). Deletions of the hrp2 and hrp3 genes result in false-negative
tests and are a threat for malaria control. A novel assay for molecular surveillance of hrp2/hrp3
deletions was developed based on droplet digital PCR (ddPCR). The assay quantifies hrp2, hrp3,
and a control gene with very high accuracy. The theoretical limit of detection was 0.33 parasites/μl.
The deletion was reliably detected in mixed infections with wild-type
and hrp2-deleted
parasites at
a density of >100 parasites/reaction. For a side-by-
side
comparison with the conventional nested
PCR (nPCR) assay, 248 samples were screened in triplicate by ddPCR and nPCR. No deletions were
observed by ddPCR, while by nPCR hrp2 deletion was observed in 8% of samples. The ddPCR assay
was applied to screen 830 samples from Kenya, Zanzibar/Tanzania, Ghana, Ethiopia, Brazil, and
Ecuador. Pronounced differences in the prevalence of deletions were observed among sites, with
more hrp3 than hrp2 deletions. In conclusion, the novel ddPCR assay minimizes the risk of false-negative
results (i.e., hrp2 deletion observed when the sample is wild type), increases sensitivity, and
greatly reduces the number of reactions that need to be run. | en_US |
| Affilliation | University of Notre Dame, Notre Dame, United States. | en_US |
| Affilliation | University of Notre Dame, Notre Dame, United States / Swiss Tropical and Public Health Institute, Allschwil, Switzerland. | en_US |
| Affilliation | Kenya Medical Research Institute-Centre for Global Health Research, Kisumu, Kenya / Department of Biochemistry and Molecular Biology, Egerton University, Nakuru, Kenya. | en_US |
| Affilliation | Kwame Nkrumah University of Science and Technology, Kumasi, Ghana. | en_US |
| Affilliation | Kwame Nkrumah University of Science and Technology, Kumasi, Ghana. | en_US |
| Affilliation | Tulane University, New Orleans, United States. | en_US |
| Affilliation | Swiss Tropical and Public Health Institute, Allschwil, Switzerland / University of Basel, Basel, Switzerland. | en_US |
| Affilliation | Swiss Tropical and Public Health Institute, Allschwil, Switzerland / University of Basel, Basel, Switzerland / Ifakara Health Institute, Dar es Salaam, United Republic of Tanzania. | en_US |
| Affilliation | Zanzibar Malaria Elimination Programme, Zanzibar, Zanzibar, United Republic of Tanzania. | en_US |
| Affilliation | Universidade de São Paulo. São Paulo, SP, Brasil. | en_US |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Doenças Parasitárias. Rio de Janeiro, RJ, Brasil. | en_US |
| Affilliation | Centro de Investigación para la Salud en América Latina, Facultad de Ciencias Exactas y Naturales, Pontificia Universidad Católica del Ecuador, Quito, Ecuador. | en_US |
| Affilliation | Department of Medical Microbiology, University of Ghana, Accra, Ghana. | en_US |
| Affilliation | Tropical and Infectious Diseases Research Center, Jimma University, Jimma, Ethiopia. | en_US |
| Affilliation | Tropical and Infectious Diseases Research Center, Jimma University, Jimma, Ethiopia. | en_US |
| Affilliation | Case Western Reserve University, Cleveland, United States. | en_US |
| Affilliation | Program in Public Health, University of California, Irvine, Irvine, United States. | en_US |
| Affilliation | University of Notre Dame, Notre Dame, United States. | en_US |
| Subject | Plasmodium falciparum | en_US |
| Subject | High-throughput | en_US |
| Subject | Hrp2 and hrp3 gene deletion typing | en_US |
| Subject | Digital PCR | en_US |
| Subject | To monitor malaria rapid | en_US |
| Subject | Diagnostic test efficacy | en_US |
