Annoura Takeshi, Chevalley Séverine, Janse Chris J, Franke-Fayard Blandine, Khan Shahid M
Center of Infectious Diseases, Leiden University Medical Center, Leiden, The Netherlands.
Methods Mol Biol. 2013;923:429-43. doi: 10.1007/978-1-62703-026-7_30.
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.
我们描述了简单且灵敏的体外和体内检测方法，以利用表达生物发光报告蛋白荧光素酶的转基因伯氏疟原虫（PbGFP-Luccon）来分析疟原虫肝期发育。在这些检测中，通过实时生物发光成像在培养物和活体小鼠中均可观察并定量肝细胞内的寄生虫发育情况。我们还描述了使用酶标仪测量发光来定量体外肝期发育的方法。基于报告基因寄生虫的肝期发育定量方法更快，并且与目前用于评估活体小鼠和培养物中肝细胞内寄生虫发育的既定定量逆转录-聚合酶链反应方法相关性非常好。
