Surveillance of Plasmodium falciparum pfcrt haplotypes in southwestern Uganda by high‐resolution melt analysis
Abstract Background Chloroquine (CQ) resistance is conferred by mutations in the Plasmodium falciparum CQ resistance transporter (pfcrt). Following CQ withdrawal for anti-malarial treatment, studies across malaria-endemic countries have shown a range of responses. In some areas, CQ sensitive parasit...
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ftdoajarticles:oai:doaj.org/article:f152dc36a8e54087990ba77a54094ca6 2023-05-15T15:17:27+02:00 Surveillance of Plasmodium falciparum pfcrt haplotypes in southwestern Uganda by high‐resolution melt analysis Kennedy Kassaza Anna C. Long Jennifer M. McDaniels Mharlove Andre Wasswa Fredrickson Dan Nyehangane Patrick Orikiriza Darwin J. Operario Joel Bazira Juliet A. Mwanga-Amumpaire Christopher C. Moore Jennifer L. Guler Yap Boum 2021-02-01T00:00:00Z https://doi.org/10.1186/s12936-021-03657-7 https://doaj.org/article/f152dc36a8e54087990ba77a54094ca6 EN eng BMC https://doi.org/10.1186/s12936-021-03657-7 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-021-03657-7 1475-2875 https://doaj.org/article/f152dc36a8e54087990ba77a54094ca6 Malaria Journal, Vol 20, Iss 1, Pp 1-10 (2021) Uganda Chloroquine Plasmodium falciparum Resistance pfcrt HRM Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2021 ftdoajarticles https://doi.org/10.1186/s12936-021-03657-7 2022-12-31T12:57:43Z Abstract Background Chloroquine (CQ) resistance is conferred by mutations in the Plasmodium falciparum CQ resistance transporter (pfcrt). Following CQ withdrawal for anti-malarial treatment, studies across malaria-endemic countries have shown a range of responses. In some areas, CQ sensitive parasites re-emerge, and in others, mutant haplotypes persist. Active surveillance of resistance mutations in clinical parasites is essential to inform treatment regimens; this effort requires fast, reliable, and cost-effective methods that work on a variety of sample types with reagents accessible in malaria-endemic countries. Methods Quantitative PCR followed by High-Resolution Melt (HRM) analysis was performed in a field setting to assess pfcrt mutations in two groups of clinical samples from Southwestern Uganda. Group 1 samples (119 in total) were collected in 2010 as predominantly Giemsa-stained slides; Group 2 samples (125 in total) were collected in 2015 as blood spots on filter paper. The Rotor-Gene Q instrument was utilized to assess the impact of different PCR-HRM reagent mixes and the detection of mixed haplotypes present in the clinical samples. Finally, the prevalence of the wild type (CVMNK) and resistant pfcrt haplotypes (CVIET and SVMNT) was evaluated in this understudied Southwestern region of Uganda. Results The sample source (i.e. Giemsa-stained slides or blood spots) and type of LCGreen-based reagent mixes did not impact the success of PCR-HRM. The detection limit of 10− 5 ng and the ability to identify mixed haplotypes as low as 10 % was similar to other HRM platforms. The CVIET haplotype predominated in the clinical samples (66 %, 162/244); however, there was a large regional variation between the sample groups (94 % CVIET in Group 1 and 44 % CVIET in Group 2). Conclusions The HRM-based method exhibits the flexibility required to conduct reliable assessment of resistance alleles from various sample types generated during the clinical management of malaria. Large regional variations in CQ resistance ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 20 1 |
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Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Uganda Chloroquine Plasmodium falciparum Resistance pfcrt HRM Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
spellingShingle |
Uganda Chloroquine Plasmodium falciparum Resistance pfcrt HRM Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Kennedy Kassaza Anna C. Long Jennifer M. McDaniels Mharlove Andre Wasswa Fredrickson Dan Nyehangane Patrick Orikiriza Darwin J. Operario Joel Bazira Juliet A. Mwanga-Amumpaire Christopher C. Moore Jennifer L. Guler Yap Boum Surveillance of Plasmodium falciparum pfcrt haplotypes in southwestern Uganda by high‐resolution melt analysis |
topic_facet |
Uganda Chloroquine Plasmodium falciparum Resistance pfcrt HRM Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Chloroquine (CQ) resistance is conferred by mutations in the Plasmodium falciparum CQ resistance transporter (pfcrt). Following CQ withdrawal for anti-malarial treatment, studies across malaria-endemic countries have shown a range of responses. In some areas, CQ sensitive parasites re-emerge, and in others, mutant haplotypes persist. Active surveillance of resistance mutations in clinical parasites is essential to inform treatment regimens; this effort requires fast, reliable, and cost-effective methods that work on a variety of sample types with reagents accessible in malaria-endemic countries. Methods Quantitative PCR followed by High-Resolution Melt (HRM) analysis was performed in a field setting to assess pfcrt mutations in two groups of clinical samples from Southwestern Uganda. Group 1 samples (119 in total) were collected in 2010 as predominantly Giemsa-stained slides; Group 2 samples (125 in total) were collected in 2015 as blood spots on filter paper. The Rotor-Gene Q instrument was utilized to assess the impact of different PCR-HRM reagent mixes and the detection of mixed haplotypes present in the clinical samples. Finally, the prevalence of the wild type (CVMNK) and resistant pfcrt haplotypes (CVIET and SVMNT) was evaluated in this understudied Southwestern region of Uganda. Results The sample source (i.e. Giemsa-stained slides or blood spots) and type of LCGreen-based reagent mixes did not impact the success of PCR-HRM. The detection limit of 10− 5 ng and the ability to identify mixed haplotypes as low as 10 % was similar to other HRM platforms. The CVIET haplotype predominated in the clinical samples (66 %, 162/244); however, there was a large regional variation between the sample groups (94 % CVIET in Group 1 and 44 % CVIET in Group 2). Conclusions The HRM-based method exhibits the flexibility required to conduct reliable assessment of resistance alleles from various sample types generated during the clinical management of malaria. Large regional variations in CQ resistance ... |
format |
Article in Journal/Newspaper |
author |
Kennedy Kassaza Anna C. Long Jennifer M. McDaniels Mharlove Andre Wasswa Fredrickson Dan Nyehangane Patrick Orikiriza Darwin J. Operario Joel Bazira Juliet A. Mwanga-Amumpaire Christopher C. Moore Jennifer L. Guler Yap Boum |
author_facet |
Kennedy Kassaza Anna C. Long Jennifer M. McDaniels Mharlove Andre Wasswa Fredrickson Dan Nyehangane Patrick Orikiriza Darwin J. Operario Joel Bazira Juliet A. Mwanga-Amumpaire Christopher C. Moore Jennifer L. Guler Yap Boum |
author_sort |
Kennedy Kassaza |
title |
Surveillance of Plasmodium falciparum pfcrt haplotypes in southwestern Uganda by high‐resolution melt analysis |
title_short |
Surveillance of Plasmodium falciparum pfcrt haplotypes in southwestern Uganda by high‐resolution melt analysis |
title_full |
Surveillance of Plasmodium falciparum pfcrt haplotypes in southwestern Uganda by high‐resolution melt analysis |
title_fullStr |
Surveillance of Plasmodium falciparum pfcrt haplotypes in southwestern Uganda by high‐resolution melt analysis |
title_full_unstemmed |
Surveillance of Plasmodium falciparum pfcrt haplotypes in southwestern Uganda by high‐resolution melt analysis |
title_sort |
surveillance of plasmodium falciparum pfcrt haplotypes in southwestern uganda by high‐resolution melt analysis |
publisher |
BMC |
publishDate |
2021 |
url |
https://doi.org/10.1186/s12936-021-03657-7 https://doaj.org/article/f152dc36a8e54087990ba77a54094ca6 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 20, Iss 1, Pp 1-10 (2021) |
op_relation |
https://doi.org/10.1186/s12936-021-03657-7 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-021-03657-7 1475-2875 https://doaj.org/article/f152dc36a8e54087990ba77a54094ca6 |
op_doi |
https://doi.org/10.1186/s12936-021-03657-7 |
container_title |
Malaria Journal |
container_volume |
20 |
container_issue |
1 |
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1766347693174554624 |