Objective

We aimed at defining the most effective routine immunoassay- or liquid chromatography-tandem mass spectrometry (LC–MS/MS)-determined steroid biomarkers for identifying non-classic adrenal hyperplasia due to 21-hydroxylase deficiency (21-NCAH) in a PCOS-like population before genotyping.

Methods

Seventy PCOS-like patients in reproductive age with immunoassay-determined follicular 17OH-progesterone (17OHP) ≥ 2.00 ng/mL underwent CYP21A2 gene analysis and _1–24ACTH test. Serum steroids were measured by immunoassays at baseline and 60 min after ACTH stimulation; basal steroid profile was measured by LC–MS/MS.

Results

Genotyping revealed 23 21-NCAH, 15 single allele heterozygous CYP21A2 mutations (21-HTZ) and 32 PCOS patients displaying similar clinical and metabolic features. Immunoassays revealed higher baseline 17OHP and testosterone, and after ACTH stimulation, higher 17OHP (17OHP₆₀) and lower cortisol, whereas LC–MS/MS revealed higher 17OHP (17OHP_LC-MS/MS), progesterone and 21-deoxycortisol and lower corticosterone in 21-NCAH compared with both 21-HTZ and PCOS patients. Steroid thresholds best discriminating 21-NCAH from 21-HTZ and PCOS were estimated, and their diagnostic accuracy in identifying 21-NCAH from PCOS was established by ROC analysis. The highest accuracy was observed for 21-deoxycortisol ≥ 0.087 ng/mL, showing 100% sensitivity, while the combination of 17OHP_LC-MS/MS ≥ 1.79 ng/mL and corticosterone ≤ 8.76 ng/mL, as well as the combination of ACTH-stimulated 17OHP ≥ 6.77 ng/mL and cortisol ≤ 240 ng/mL by immunoassay, showed 100% specificity.

Conclusions

LC–MS/MS measurement of basal follicular 21-deoxycortisol, 17OHP and corticosterone seems the most convenient method for diagnosing 21-NCAH in a population of PCOS with a positive first level screening, providing high accuracy and reducing the need for ACTH stimulation test.

中文翻译：

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类固醇生物标志物，用于在可疑水平为17OH-孕酮的PCOS人群中鉴定由于21-羟化酶缺乏症引起的非经典肾上腺增生。

目的

我们旨在定义最有效的常规免疫测定或液相色谱-串联质谱（LC-MS / MS）测定的类固醇生物标记物，以鉴定PCOS-中由于21-羟化酶缺乏症（21-NCAH）引起的非经典肾上腺增生。就像基因分型前的人口

方法

通过免疫测定法测定的卵泡17OH-孕酮（17OHP）≥2.00 ng / mL的70名育龄PCOS样患者接受了CYP21A2基因分析和_1-24 ACTH测试。在基线和促肾上腺皮质激素刺激后60分钟，通过免疫测定法测定血清类固醇。通过LC-MS / MS测量基础类固醇激素谱。

结果

基因分型显示23例21-NCAH，15例单等位基因杂合CYP21A2突变（21-HTZ）和32例PCOS患者表现出相似的临床和代谢特征。免疫测定显示较高的基线17OHP和睾丸激素，在ACTH刺激后，较高的17OHP（17OHP ₆₀）和较低的皮质醇，而LC–MS / MS显示较高的17OHP（17OHP _{LC-MS / MS}），与21-HTZ和PCOS患者相比，21-NCAH中的孕酮和21-脱氧皮质醇和较低的皮质酮。估计了最佳区分21-NCAH与21-HTAH和PCOS的类固醇阈值，并通过ROC分析确定了它们在从PCOS识别21-NCAH中的诊断准确性。对于21-脱氧皮质醇≥0.087 ng / mL观察到最高的准确性，显示出100％的敏感性，而17OHP _{LC-MS /} MS≥1.79  ng / mL的组合和皮质酮≤8.76 ng / mL的组合以及ACTH的组合通过免疫测定刺激的17OHP≥6.77 ng / mL和皮质醇≤240 ng / mL，表现出100％的特异性。

结论

LC-MS / MS测定基础滤泡性21-脱氧皮质醇，17OHP和皮质酮似乎是诊断PCOS人群中21-NCAH的最便捷的方法，具有积极的一级筛查，可提供高精度并减少对ACTH刺激试验的需要。