Journal
JOURNAL OF INFECTIOUS DISEASES
Volume 225, Issue 9, Pages 1621-1625Publisher
OXFORD UNIV PRESS INC
DOI: 10.1093/infdis/jiab431
Keywords
flow analyses; Plasmodium; RNA-FISH
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Funding
- French Parasitology Consortium ParaFrap [ANR-11-LABX0024]
- Fondation Pasteur Suisse
- Pasteur Cantarini-Roux postdoc fellowship
- Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP), Brazil [2016/18740-9]
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The new Flow-FISH method combines RNA detection and cell staining techniques to accurately quantify different stages of malaria parasites, suitable for gene expression studies and analysis of liver-stage sporozoites.
Thanks to the combination of Plasmodium-specific RNA detection and surface and nucleus staining, the new Flow-FISH method allowed us to characterize and quantify asexual and sexual blood-stage parasite from P vivax clinical samples as well as liver-stage parasites. We adapted the RNA FISH Stellaris method to specifically detect the expression of Plasmodium genes by flow cytometry and ImageStream (Flow-FISH). This new method accurately quantified the erythrocytic forms of (1) Plasmodium falciparum and Plasmodium vivax and (2) the sexual stages of P vivax from patient isolates. ImageStream analysis of liver stage sporozoites using a combination of surface circumsporozoite protein (CSP), deoxyribonucleic acid, and 18S RNA labeling proved that the new Flow-FISH is suitable for gene expression studies of transmission stages. This powerful multiparametric single-cell method offers a platform of choice for both applied and fundamental research on the biology of malaria parasites.
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