A semi-automated method for counting fluorescent malaria oocysts increases the throughput of transmission blocking studies
Abstract Background Malaria transmission is now recognized as a key target for intervention. Evaluation of the Plasmodium oocyst burden in the midguts of Anopheles spp . is important for many of assays investigating transmission. However, current assays are very time-consuming, manually demanding an...
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ftdoajarticles:oai:doaj.org/article:9e6c0f60f2af4cf9b4232966b97797b1 2023-05-15T15:09:56+02:00 A semi-automated method for counting fluorescent malaria oocysts increases the throughput of transmission blocking studies Sinden Robert E Delves Michael J 2010-01-01T00:00:00Z https://doi.org/10.1186/1475-2875-9-35 https://doaj.org/article/9e6c0f60f2af4cf9b4232966b97797b1 EN eng BMC http://www.malariajournal.com/content/9/1/35 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-9-35 1475-2875 https://doaj.org/article/9e6c0f60f2af4cf9b4232966b97797b1 Malaria Journal, Vol 9, Iss 1, p 35 (2010) Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2010 ftdoajarticles https://doi.org/10.1186/1475-2875-9-35 2022-12-31T13:00:14Z Abstract Background Malaria transmission is now recognized as a key target for intervention. Evaluation of the Plasmodium oocyst burden in the midguts of Anopheles spp . is important for many of assays investigating transmission. However, current assays are very time-consuming, manually demanding and patently subject to observer-observer variation. Methods This report presents the development of a method to rapidly, accurately and consistently determine oocyst burdens on mosquito midguts using GFP-expressing Plasmodium berghei and a custom-written macro for ImageJ. The counting macro was optimized and found to be fit-for-purpose by performing gametocyte membrane feeds with parasite infected blood. Dissected midguts were counted both manually and using the automated macro, then compared. The optimized settings for the macro were then validated by using it to determine the transmission blocking efficacies of two anti-malarial compounds - dehydroepiandrosterone sulphate and lumefantrine, in comparison to manually determined analysis of the same experiment. Results Concurrence of manual and macro counts was very high (R 2 = 0.973) and reproducible. Estimated transmission blocking efficacies between manual and automated analysis were highly concordant, indicating that dehydroepiandrosterone sulphate has little or no transmission blocking potential, whilst lumefantrine strongly inhibits sporogony. Conclusion Recognizing a potential five-fold increase in throughput, the resulting reduction in personnel costs, and the absence of inter-operator/laboratory variation possible with this approach, this counting macro may be a benefit to the malaria community. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 9 1 |
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Directory of Open Access Journals: DOAJ Articles |
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English |
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Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
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Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Sinden Robert E Delves Michael J A semi-automated method for counting fluorescent malaria oocysts increases the throughput of transmission blocking studies |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Malaria transmission is now recognized as a key target for intervention. Evaluation of the Plasmodium oocyst burden in the midguts of Anopheles spp . is important for many of assays investigating transmission. However, current assays are very time-consuming, manually demanding and patently subject to observer-observer variation. Methods This report presents the development of a method to rapidly, accurately and consistently determine oocyst burdens on mosquito midguts using GFP-expressing Plasmodium berghei and a custom-written macro for ImageJ. The counting macro was optimized and found to be fit-for-purpose by performing gametocyte membrane feeds with parasite infected blood. Dissected midguts were counted both manually and using the automated macro, then compared. The optimized settings for the macro were then validated by using it to determine the transmission blocking efficacies of two anti-malarial compounds - dehydroepiandrosterone sulphate and lumefantrine, in comparison to manually determined analysis of the same experiment. Results Concurrence of manual and macro counts was very high (R 2 = 0.973) and reproducible. Estimated transmission blocking efficacies between manual and automated analysis were highly concordant, indicating that dehydroepiandrosterone sulphate has little or no transmission blocking potential, whilst lumefantrine strongly inhibits sporogony. Conclusion Recognizing a potential five-fold increase in throughput, the resulting reduction in personnel costs, and the absence of inter-operator/laboratory variation possible with this approach, this counting macro may be a benefit to the malaria community. |
format |
Article in Journal/Newspaper |
author |
Sinden Robert E Delves Michael J |
author_facet |
Sinden Robert E Delves Michael J |
author_sort |
Sinden Robert E |
title |
A semi-automated method for counting fluorescent malaria oocysts increases the throughput of transmission blocking studies |
title_short |
A semi-automated method for counting fluorescent malaria oocysts increases the throughput of transmission blocking studies |
title_full |
A semi-automated method for counting fluorescent malaria oocysts increases the throughput of transmission blocking studies |
title_fullStr |
A semi-automated method for counting fluorescent malaria oocysts increases the throughput of transmission blocking studies |
title_full_unstemmed |
A semi-automated method for counting fluorescent malaria oocysts increases the throughput of transmission blocking studies |
title_sort |
semi-automated method for counting fluorescent malaria oocysts increases the throughput of transmission blocking studies |
publisher |
BMC |
publishDate |
2010 |
url |
https://doi.org/10.1186/1475-2875-9-35 https://doaj.org/article/9e6c0f60f2af4cf9b4232966b97797b1 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 9, Iss 1, p 35 (2010) |
op_relation |
http://www.malariajournal.com/content/9/1/35 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-9-35 1475-2875 https://doaj.org/article/9e6c0f60f2af4cf9b4232966b97797b1 |
op_doi |
https://doi.org/10.1186/1475-2875-9-35 |
container_title |
Malaria Journal |
container_volume |
9 |
container_issue |
1 |
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1766341028554473472 |