A simple monochromatic flow cytometric assay for assessment of intraerythrocytic development of Plasmodium falciparum
Abstract Background Gold standard microscopic examination of Plasmodium falciparum intraerythrocytic stage remains an important process for staging and enumerating parasitized erythrocytes in culture; however, microscopy is laborious and its accuracy is dependent upon the skill of the examiner. Meth...
Published in: | Malaria Journal |
---|---|
Main Authors: | , , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
BMC
2020
|
Subjects: | |
Online Access: | https://doi.org/10.1186/s12936-020-03156-1 https://doaj.org/article/f0b4de191bc44a3799fce8e695bead2e |
id |
ftdoajarticles:oai:doaj.org/article:f0b4de191bc44a3799fce8e695bead2e |
---|---|
record_format |
openpolar |
spelling |
ftdoajarticles:oai:doaj.org/article:f0b4de191bc44a3799fce8e695bead2e 2023-05-15T15:08:33+02:00 A simple monochromatic flow cytometric assay for assessment of intraerythrocytic development of Plasmodium falciparum Kasem Kulkeaw Nutpakal Ketprasit Anchalee Tungtrongchitr Duangdao Palasuwan 2020-02-01T00:00:00Z https://doi.org/10.1186/s12936-020-03156-1 https://doaj.org/article/f0b4de191bc44a3799fce8e695bead2e EN eng BMC http://link.springer.com/article/10.1186/s12936-020-03156-1 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-020-03156-1 1475-2875 https://doaj.org/article/f0b4de191bc44a3799fce8e695bead2e Malaria Journal, Vol 19, Iss 1, Pp 1-13 (2020) Malaria parasite development Monochromatic flow cytometric assay Plasmodium falciparum VSG Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2020 ftdoajarticles https://doi.org/10.1186/s12936-020-03156-1 2022-12-31T10:41:16Z Abstract Background Gold standard microscopic examination of Plasmodium falciparum intraerythrocytic stage remains an important process for staging and enumerating parasitized erythrocytes in culture; however, microscopy is laborious and its accuracy is dependent upon the skill of the examiner. Methods In this study, ViSafe Green (VSG), which is a nucleic acid-binding fluorescent dye, was used for assessing in vitro development of P. falciparum using flow cytometry. Results Fluorescence intensity of VSG was found to depend on the developmental stage of parasites. Specifically, multiple-nuclei-containing schizonts were observed in the VSGhigh population, and growing trophozoites and ring-shaped forms were observed in the VSGintermediate and VSGlow populations. The efficacy of VSG-based assay was found to be comparable to the microscopic examination method, and it demonstrated an ability to detect as low as 0.001% of the parasitaemia estimated by Giemsa staining. Moreover, when applying VSG for anti-malarial drug test, it was able to observe the growth inhibitory effect of dihydroartemisinin, the front-line drug for malaria therapy. Conclusions Taken together, the results of this study suggest the VSG-based flow cytometric assay to be a simple and reliable assay for assessing P. falciparum malaria development in vitro. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 19 1 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Malaria parasite development Monochromatic flow cytometric assay Plasmodium falciparum VSG Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
spellingShingle |
Malaria parasite development Monochromatic flow cytometric assay Plasmodium falciparum VSG Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Kasem Kulkeaw Nutpakal Ketprasit Anchalee Tungtrongchitr Duangdao Palasuwan A simple monochromatic flow cytometric assay for assessment of intraerythrocytic development of Plasmodium falciparum |
topic_facet |
Malaria parasite development Monochromatic flow cytometric assay Plasmodium falciparum VSG Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Gold standard microscopic examination of Plasmodium falciparum intraerythrocytic stage remains an important process for staging and enumerating parasitized erythrocytes in culture; however, microscopy is laborious and its accuracy is dependent upon the skill of the examiner. Methods In this study, ViSafe Green (VSG), which is a nucleic acid-binding fluorescent dye, was used for assessing in vitro development of P. falciparum using flow cytometry. Results Fluorescence intensity of VSG was found to depend on the developmental stage of parasites. Specifically, multiple-nuclei-containing schizonts were observed in the VSGhigh population, and growing trophozoites and ring-shaped forms were observed in the VSGintermediate and VSGlow populations. The efficacy of VSG-based assay was found to be comparable to the microscopic examination method, and it demonstrated an ability to detect as low as 0.001% of the parasitaemia estimated by Giemsa staining. Moreover, when applying VSG for anti-malarial drug test, it was able to observe the growth inhibitory effect of dihydroartemisinin, the front-line drug for malaria therapy. Conclusions Taken together, the results of this study suggest the VSG-based flow cytometric assay to be a simple and reliable assay for assessing P. falciparum malaria development in vitro. |
format |
Article in Journal/Newspaper |
author |
Kasem Kulkeaw Nutpakal Ketprasit Anchalee Tungtrongchitr Duangdao Palasuwan |
author_facet |
Kasem Kulkeaw Nutpakal Ketprasit Anchalee Tungtrongchitr Duangdao Palasuwan |
author_sort |
Kasem Kulkeaw |
title |
A simple monochromatic flow cytometric assay for assessment of intraerythrocytic development of Plasmodium falciparum |
title_short |
A simple monochromatic flow cytometric assay for assessment of intraerythrocytic development of Plasmodium falciparum |
title_full |
A simple monochromatic flow cytometric assay for assessment of intraerythrocytic development of Plasmodium falciparum |
title_fullStr |
A simple monochromatic flow cytometric assay for assessment of intraerythrocytic development of Plasmodium falciparum |
title_full_unstemmed |
A simple monochromatic flow cytometric assay for assessment of intraerythrocytic development of Plasmodium falciparum |
title_sort |
simple monochromatic flow cytometric assay for assessment of intraerythrocytic development of plasmodium falciparum |
publisher |
BMC |
publishDate |
2020 |
url |
https://doi.org/10.1186/s12936-020-03156-1 https://doaj.org/article/f0b4de191bc44a3799fce8e695bead2e |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 19, Iss 1, Pp 1-13 (2020) |
op_relation |
http://link.springer.com/article/10.1186/s12936-020-03156-1 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-020-03156-1 1475-2875 https://doaj.org/article/f0b4de191bc44a3799fce8e695bead2e |
op_doi |
https://doi.org/10.1186/s12936-020-03156-1 |
container_title |
Malaria Journal |
container_volume |
19 |
container_issue |
1 |
_version_ |
1766339893589442560 |