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Flowcytometric and ImageStream Rna-Fish Gene Expression, Quantification and Phenotypic Characterization of Blood Sporozoites and Sporozoites From Human Malaria Species

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Autor(es):
Luiza-Batista, Camilla ; Nardella, Flore ; Thiberge, Sabine ; Serra-Hassoun, Malika ; Ferreira, Marcelo U. ; Scherf, Artur ; Garcia, Sylvie
Número total de Autores: 7
Tipo de documento: Artigo Científico
Fonte: Journal of Infectious Diseases; v. 225, n. 9, p. 5-pg., 2021-08-28.
Resumo

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. (AU)

Processo FAPESP: 16/18740-9 - Bases científicas para a eliminação da malária residual na Amazônia Brasileira
Beneficiário:Marcelo Urbano Ferreira
Modalidade de apoio: Auxílio à Pesquisa - Temático