背景:在数量/体积有限的临床样本中快速准确地检测17α-羟孕酮(17-OHP)和相关类固醇的方法对于临床上精确诊断新生儿先天性肾上腺增生(CAH)及其亚型具有重要意义。
方法:16种类固醇(17-OHP,雄烯二酮,皮质醇,四氢-11-脱氧皮质醇,孕烯醇酮,黄体酮,11-脱氧皮质酮,皮质酮,21-脱氧皮质醇,11-脱氧皮质醇,脱氢表雄酮,睾丸激素,醛固酮,17α-羟基孕烯醇酮,双氢睾酮和18-羟基皮质酮)包括在高通量微孔超高效液相色谱-串联质谱组中。从126名正常受试者和65名包括不同CAH亚型的患者收集样品。
结果:该方法在线性方面具有令人满意的分析性能,重复性,回收率和检测限。通过量化正常婴儿中检测到的类固醇水平来建立16种类固醇的参考间隔。通过区分正常婴儿和CAH婴儿的类固醇代谢特征来测试该方法的适用性。以及具有不同CAH亚型的婴儿之间。17-OHP的相关性,21-脱氧皮质醇,并证明了17-OHP/11-脱氧皮质醇用于21-羟化酶缺乏症筛查。11-脱氧皮质酮的水平,11-脱氧皮质醇,孕酮和雄烯二酮可用于诊断CAH的不同罕见亚型。
结论:这项研究为数量/体积有限的临床样本提供了一种高效的类固醇分析策略,并具有临床诊断CAH的巨大潜力。
BACKGROUND: A rapid and accurate measurement approach for 17α-hydroxyprogesterone (17-OHP) and related steroids in amount/volume-limited clinic samples is of importance for precise newborn diagnosis of congenital adrenal hyperplasia (CAH) and its subtypes in clinic.
METHODS: Sixteen steroids (17-OHP, androstenedione, cortisol, tetrahydro-11-deoxycortisol, pregnenolone, progesterone, 11-deoxycorticosterone, corticosterone, 21-deoxycortisol, 11-deoxycortisol, dehydroepiandrosterone, testosterone, aldosterone, 17α-hydroxypregnenolone, dihydrotestosterone and 18-hydroxycorticosterone) were included in the panel of high-throughput microbore ultra-performance liquid chromatography-tandem mass spectrometry. Samples were collected from 126 normal subjects and 65 patients including different subtypes of CAH.
RESULTS: The method was validated with satisfactory analytical performance in linearity, repeatability, recovery and limit of detection. Reference intervals for 16 steroids were established by quantifying the level of steroids detected in normal infants. The applicability of the method was tested by differentiating steroid metabolic characteristics between normal infants and infants with CAH, as well as between infants with different CAH subtypes. The relevance of 17-OHP, 21-deoxycortisol, and 17-OHP/11-deoxycortisol for 21-hydroxylase deficiency screening was demonstrated. The level of 11-deoxycorticosterone, 11-deoxycortisol, progesterone and androstenedione can be used for the diagnosis of different rare subtypes of CAH.
CONCLUSIONS: This study provides a strategy for highly efficient steroid analysis of amount/volume-limited clinic samples and holds great potential for clinical diagnosis of CAH.