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Adaptation of a Gaussia princeps Luciferase reporter system in Candida albicans for in vivo detection in the Galleria mellonella infection model

机译:适应性高斯王子荧光素酶报告系统在白色念珠菌中用于在梅勒埃勒菌感染模型中进行体内检测

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For the past 10 years, mini-host models and in particular the greater wax moth Galleria mellonella have tended to become a surrogate for murine models of fungal infection mainly due to cost, ethical constraints and ease of use. Thus, methods to better assess the fungal pathogenesis in G. mellonella need to be developed. In this study, we implemented the detection of Candida albicans cells expressing the Gaussia princeps luciferase in its cell wall in infected larvae of G. mellonella. We demonstrated that detection and quantification of luminescence in the pulp of infected larvae is a reliable method to perform drug efficacy and C. albicans virulence assays as compared to fungal burden assay. Since the linearity of the bioluminescent signal, as compared to the CFU counts, has a correlation of R-2 = 0.62 and that this method is twice faster and less labor intensive than classical fungal burden assays, it could be applied to large scale studies. We next visualized and followed C. albicans infection in living G. mellonella larvae using a non-toxic and water-soluble coelenterazine formulation and a CCD camera that is commonly used for chemoluminescence signal detection. This work allowed us to follow for the first time C. albicans course of infection in G. mellonella during 4 days.
机译:在过去的十年中,主要由于成本,道德约束和易用性,微型宿主模型,尤其是较大的蜡蛾梅花菌已趋于成为鼠类真菌感染模型的替代品。因此,需要开发一种方法来更好地评估苜蓿根瘤菌的真菌发病机理。在这项研究中,我们实施了检测白色念珠菌细胞在感染的G. mellonella幼虫的细胞壁中表达高斯princeps荧光素酶的细胞。我们证明了检测和定量感染的幼虫果肉中的发光是执行药物功效和白色念珠菌毒力测定的可靠方法,与真菌负荷测定相比。由于与CFU计数相比,生物发光信号的线性度具有R-2 = 0.62的相关性,并且该方法比传统的真菌负荷测定法快两倍,且劳动强度较小,因此可以用于大规模研究。接下来,我们使用无毒且水溶性的腔肠素制剂和通常用于化学发光信号检测的CCD摄像机,对活的G. mellonella幼虫中的白色念珠菌感染进行了观察和跟踪。这项工作使我们能够在4天之内首次追踪白色念珠菌感染梅花菌的过程。

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