Quantitative Analysis of Plasmodium berghei Liver Stages by Bioluminescence Imaging
We describe simple and sensitive in vitro and in vivo assays to analyze Plasmodium liver stage development using transgenic P. berghei parasites (PbGFP-Luccon), which express the bioluminescent reporter protein, luciferase. In these assays, parasite development in hepatocytes is visualized and quantified by real-time bioluminescence imaging both in culture and in live mice. We also describe quantification of in vitro liver-stage development by measuring luminescence using a microplate reader. Reporter-parasite based quantification of liver-stage development is faster and correlates very well with established quantitative RT-PCR methods currently used to assess parasite development inside hepatocytes, both in live mice and in culture.
Key wordsMalaria Plasmodium berghei Sporozoites Preerythrocytic stages Liver Hepatocytes Luciferase Luminescence In vivo imaging Drug screening Mice
TA, SC, and SMK received financial support of Top Institute Pharma (The Netherlands) project: T4-102.
- 13.Sinden RE (1997) Infection of mosquitoes with rodent malaria. in The molecular biology of insect disease vectors. eds Crampton J. M., Beard C. B., Louis C. (Chapman & Hall, London, United Kingdom), pp 261–267.Google Scholar