Arias M. H., Deharo Eric, Valentin A., Garavito G. (2017). Adaptation and optimization of a fluorescence-based assay for in vivo antimalarial drug screening. Parasitology Research, 116 (7), p. 1955-1962. ISSN 0932-0113.
Titre du document
Adaptation and optimization of a fluorescence-based assay for in vivo antimalarial drug screening
Arias M. H., Deharo Eric, Valentin A., Garavito G.
Source
Parasitology Research, 2017,
116 (7), p. 1955-1962 ISSN 0932-0113
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.
