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Tests in vitro de l'action inhibitrice de la croissance de P. falciparum par les antipaludiques
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- 25. 25 Dye-Based incorporation for antimalarial drug testing • SYBR Green I – Smilkstein et al, 2004 (AAC, May 2004) • In vitro cultivated parasites • Compared SYBR green to [3H]ethanolamine incorporation • Linear relationship between parasitemia and measured fluorescence (0%-13.5%) • Similar EC50 compared to [3H]ethanolamine incorporation – Bennett et al, 2004 (AAC, May 2004) • In vitro cultivated parasites • Similar SYBR green I staining characteristics seen with various stages of parasites • Similar IC50 with published methods (3H) • Picogreen – Corbett et al, 2004 (AJTMH, 2004) • In vitro cultivated parasites • Compared Picogreen to 3H hypoxanthine incorporation • Linear relation between fluorescence and % parasitemia (0.1%-15%)
- 26. 26 incubate 1 h read transfer 100ul add lysis buffer with SYBR green Freeze plates Incubate culture (72h) Plate set-up SYBR Green HRPII ELISA Coat plates Incubate 1 h, wash incubate 1 h, wash incubate 1 h, wash read Add developer Add detector antibody Add anti-HRPII Add sample Freeze plate Incubate culture (72h) Plate set-up
- 27. 27 SYBR Green vs HRPII 3D7 cultured parasites EC5050value(ng/ml) 0 2 4 6 8 10 12 CQ Q MQ ART SYBR HRPII
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- 38. 38 Les différents types de puces disponibles à l’IPC : 1 ) B . ( B ) / < < ( U : 1 ) ( . ) 1 (?
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