%0 Journal Article
%9 ACL : Articles dans des revues avec comité de lecture répertoriées par l'AERES
%A Arias, M. H.
%A Deharo, Eric
%A Valentin, A.
%A Garavito, G.
%T Adaptation and optimization of a fluorescence-based assay for in vivo antimalarial drug screening
%D 2017
%L fdi:010070246
%G ENG
%J Parasitology Research
%@ 0932-0113
%K Malaria ; Pharmacology ; Plasmodium berghei ; SYBR Green I
%M ISI:000403940600022
%N 7
%P 1955-1962
%R 10.1007/s00436-017-5477-z
%U https://www.documentation.ird.fr/hor/fdi:010070246
%> https://www.documentation.ird.fr/intranet/publi/2017/07/010070246.pdf
%V 116
%W Horizon (IRD)
%X The in vivo efficacy of potential antimalarials is usually evaluated by direct microscopic determination of the parasitaemia of Plasmodium-infected mice on Giemsa-stained blood smears. This process is time-consuming, requires experienced technicians and is not automatable. Therefore, we optimized a SYBR Green I (SYBRG I) fluorescence-based assay to fluorometers commonly available in many research laboratories. This technique was originally developed to assess parasitaemia in humans by cytometry. We defined optimal conditions with Plasmodium berghei-infected mice, standard lysis buffer (Tris, EDTA, saponin and Triton), whole blood cells and 2 h staining incubation with SYBRG I 2X. The fluorescence background generated by uninfected whole blood cells was low (around 4.6%), and the linearity high (r(2) = 0.96), with parasitaemia ranging from 1.4 to 60%. The Bland-Altman plot showed a strong correlation between SYBRG I and Giemsa gold standard method; Z'-factor was >0.5. These findings suggest that our fluorescence-based assay is suitable for in vivo antimalarial drug assessment in a malaria murine model. It can help to overcome the human bias found with microscopic techniques.
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