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Feasibility of community at-home dried blood spot collection combined with pooled reverse transcription PCR as a viable and convenient method for malaria epidemiology studies

BACKGROUND: Many Plasmodium infections in endemic regions exist at densities below the limit of detection of standard diagnostic tools. These infections threaten control efforts and may impact vaccine and therapeutic drug studies. Simple, cost-effective methods are needed to study the natural histor...

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Autores principales: Hergott, Dianna E. B., Owalla, Tonny J., Balkus, Jennifer E., Apio, Bernadette, Lema, Jimmy, Cemeri, Barbara, Akileng, Andrew, Seilie, Annette M., Chavtur, Chris, Staubus, Weston, Chang, Ming, Egwang, Thomas G., Murphy, Sean C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9284728/
https://www.ncbi.nlm.nih.gov/pubmed/35836179
http://dx.doi.org/10.1186/s12936-022-04239-x
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author Hergott, Dianna E. B.
Owalla, Tonny J.
Balkus, Jennifer E.
Apio, Bernadette
Lema, Jimmy
Cemeri, Barbara
Akileng, Andrew
Seilie, Annette M.
Chavtur, Chris
Staubus, Weston
Chang, Ming
Egwang, Thomas G.
Murphy, Sean C.
author_facet Hergott, Dianna E. B.
Owalla, Tonny J.
Balkus, Jennifer E.
Apio, Bernadette
Lema, Jimmy
Cemeri, Barbara
Akileng, Andrew
Seilie, Annette M.
Chavtur, Chris
Staubus, Weston
Chang, Ming
Egwang, Thomas G.
Murphy, Sean C.
author_sort Hergott, Dianna E. B.
collection PubMed
description BACKGROUND: Many Plasmodium infections in endemic regions exist at densities below the limit of detection of standard diagnostic tools. These infections threaten control efforts and may impact vaccine and therapeutic drug studies. Simple, cost-effective methods are needed to study the natural history of asymptomatic submicroscopic parasitaemia. Self-collected dried blood spots (DBS) analysed using pooled and individual quantitative reverse transcription polymerase chain reaction (qRT-PCR) provide such a solution. Here, the feasibility and acceptability of daily at-home DBS collections for qRT-PCR was studied to better understand low-density infections. METHODS: Rapid diagnostic test (RDT)-negative individuals in Katakwi District, northeastern Uganda, were recruited between April and May 2021. Venous blood samples and clinic-collected DBS were taken at enrollment and at four weekly clinic visits. Participants were trained in DBS collection and asked to collect six DBS weekly between clinic visits. Opinions about the collection process were solicited using daily Diary Cards and a Likert scale survey at the final study visit. Venous blood and DBS were analysed by Plasmodium 18S rRNA qRT-PCR. The number of participants completing the study, total DBS collected, and opinions of the process were analysed to determine compliance and acceptability. The human internal control mRNA and Plasmodium 18S rRNA were evaluated for at-home vs. clinic-collected DBS and venous blood to assess quality and accuracy of at-home collected samples. RESULTS: One-hundred two adults and 29 children were enrolled, and 95 and 26 completed the study, respectively. Three individuals withdrew due to pain or inconvenience of procedures. Overall, 96% of participants collected ≥ 16 of 24 at-home DBS, and 87% of DBS contained ≥ 40 µL of blood. The procedure was well tolerated and viewed favourably by participants. At-home collected DBS were acceptable for qRT-PCR and showed less than a one qRT-PCR cycle threshold shift in the human control mRNA compared to clinic-collected DBS. Correlation between Plasmodium falciparum 18S rRNA from paired whole blood and DBS was high (R = 0.93). CONCLUSIONS: At-home DBS collection is a feasible, acceptable, and robust method to obtain blood to evaluate the natural history of low-density Plasmodium infections by qRT-PCR. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12936-022-04239-x.
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spelling pubmed-92847282022-07-16 Feasibility of community at-home dried blood spot collection combined with pooled reverse transcription PCR as a viable and convenient method for malaria epidemiology studies Hergott, Dianna E. B. Owalla, Tonny J. Balkus, Jennifer E. Apio, Bernadette Lema, Jimmy Cemeri, Barbara Akileng, Andrew Seilie, Annette M. Chavtur, Chris Staubus, Weston Chang, Ming Egwang, Thomas G. Murphy, Sean C. Malar J Research BACKGROUND: Many Plasmodium infections in endemic regions exist at densities below the limit of detection of standard diagnostic tools. These infections threaten control efforts and may impact vaccine and therapeutic drug studies. Simple, cost-effective methods are needed to study the natural history of asymptomatic submicroscopic parasitaemia. Self-collected dried blood spots (DBS) analysed using pooled and individual quantitative reverse transcription polymerase chain reaction (qRT-PCR) provide such a solution. Here, the feasibility and acceptability of daily at-home DBS collections for qRT-PCR was studied to better understand low-density infections. METHODS: Rapid diagnostic test (RDT)-negative individuals in Katakwi District, northeastern Uganda, were recruited between April and May 2021. Venous blood samples and clinic-collected DBS were taken at enrollment and at four weekly clinic visits. Participants were trained in DBS collection and asked to collect six DBS weekly between clinic visits. Opinions about the collection process were solicited using daily Diary Cards and a Likert scale survey at the final study visit. Venous blood and DBS were analysed by Plasmodium 18S rRNA qRT-PCR. The number of participants completing the study, total DBS collected, and opinions of the process were analysed to determine compliance and acceptability. The human internal control mRNA and Plasmodium 18S rRNA were evaluated for at-home vs. clinic-collected DBS and venous blood to assess quality and accuracy of at-home collected samples. RESULTS: One-hundred two adults and 29 children were enrolled, and 95 and 26 completed the study, respectively. Three individuals withdrew due to pain or inconvenience of procedures. Overall, 96% of participants collected ≥ 16 of 24 at-home DBS, and 87% of DBS contained ≥ 40 µL of blood. The procedure was well tolerated and viewed favourably by participants. At-home collected DBS were acceptable for qRT-PCR and showed less than a one qRT-PCR cycle threshold shift in the human control mRNA compared to clinic-collected DBS. Correlation between Plasmodium falciparum 18S rRNA from paired whole blood and DBS was high (R = 0.93). CONCLUSIONS: At-home DBS collection is a feasible, acceptable, and robust method to obtain blood to evaluate the natural history of low-density Plasmodium infections by qRT-PCR. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12936-022-04239-x. BioMed Central 2022-07-14 /pmc/articles/PMC9284728/ /pubmed/35836179 http://dx.doi.org/10.1186/s12936-022-04239-x Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Hergott, Dianna E. B.
Owalla, Tonny J.
Balkus, Jennifer E.
Apio, Bernadette
Lema, Jimmy
Cemeri, Barbara
Akileng, Andrew
Seilie, Annette M.
Chavtur, Chris
Staubus, Weston
Chang, Ming
Egwang, Thomas G.
Murphy, Sean C.
Feasibility of community at-home dried blood spot collection combined with pooled reverse transcription PCR as a viable and convenient method for malaria epidemiology studies
title Feasibility of community at-home dried blood spot collection combined with pooled reverse transcription PCR as a viable and convenient method for malaria epidemiology studies
title_full Feasibility of community at-home dried blood spot collection combined with pooled reverse transcription PCR as a viable and convenient method for malaria epidemiology studies
title_fullStr Feasibility of community at-home dried blood spot collection combined with pooled reverse transcription PCR as a viable and convenient method for malaria epidemiology studies
title_full_unstemmed Feasibility of community at-home dried blood spot collection combined with pooled reverse transcription PCR as a viable and convenient method for malaria epidemiology studies
title_short Feasibility of community at-home dried blood spot collection combined with pooled reverse transcription PCR as a viable and convenient method for malaria epidemiology studies
title_sort feasibility of community at-home dried blood spot collection combined with pooled reverse transcription pcr as a viable and convenient method for malaria epidemiology studies
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9284728/
https://www.ncbi.nlm.nih.gov/pubmed/35836179
http://dx.doi.org/10.1186/s12936-022-04239-x
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