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

Author(s)

Luiza-Batista, Camilla ; Nardella, Flore ; Thiberge, Sabine ; Serra-Hassoun, Malika ; Ferreira, Marcelo U. ; Scherf, Artur ; Garcia, Sylvie

• Document type: Journal article
• Source: Journal of Infectious Diseases; v. 225, n. 9, p. 5-pg., 2021-08-28.

Abstract

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)

• FAPESP's process: 16/18740-9 - Scientific bases for residual malaria elimination in the Brazilian Amazon
• Grantee: Marcelo Urbano Ferreira
• Support Opportunities: Research Projects - Thematic Grants