Bennett Michael J, Sheng Feng, Saada Ann
Department of Pathology & Laboratory Medicine, University of Pennsylvania Perelman School of Medicine, Metabolic Disease Laboratory, The Children's Hospital of Philadelphia, Philadelphia, PA, United States.
Department of Pathology & Laboratory Medicine, University of Pennsylvania Perelman School of Medicine, Metabolic Disease Laboratory, The Children's Hospital of Philadelphia, Philadelphia, PA, United States.
Methods Cell Biol. 2020;155:83-120. doi: 10.1016/bs.mcb.2019.11.021. Epub 2020 Jan 21.
This chapter focuses on the methods to measure unique metabolites, specific enzymes, and metabolic flux in fatty acid β-oxidation, and on biochemical assays of tricarboxylic acid (TCA) cycle enzymes and the pyruvate dehydrogenase complex. These assays play an important role in the diagnosis of genetic diseases, newborn screening, and in cancer and metabolism research. The rationale, protocol, pros and cons, and alternative methods are discussed. Nevertheless, each laboratory should adapt the preferred method optimizing sample preparation and assay conditions for linearity and a low signal-to-noise ratio. The reader is also referred to the additional literature citing methods and clinical descriptions of the various diseases.
本章重点介绍测量脂肪酸β-氧化中独特代谢物、特定酶和代谢通量的方法，以及三羧酸（TCA）循环酶和丙酮酸脱氢酶复合体的生化检测方法。这些检测在遗传疾病诊断、新生儿筛查以及癌症和代谢研究中发挥着重要作用。文中讨论了其基本原理、实验方案、优缺点及替代方法。然而，每个实验室都应采用首选方法，优化样品制备和检测条件，以实现线性和低信噪比。读者还可参考引用各种疾病的方法和临床描述的其他文献。
