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基于气相色谱-质谱联用技术分析尿石症患者血清与尿液氨基酸代谢轮廓

Serum and urinary amino acid profiling analysis for urinary stone disease using GC-MS

摘要:

目的:分析尿石症患者血清与尿液的氨基酸代谢轮廓,寻找疾病相关的差异生物标志物,为临床早期筛查诊断提供可靠依据。方法:病例对照研究。收集2015年2月至2017年10月福建医科大学附属第一医院泌尿外科确诊的74例尿石症患者(年龄20~82岁,男41例,女33例)和同期健康体检中心的35名健康对照者(年龄22~80岁,男20名,女15名)的血液与尿液样本。采用基于GC-MS的代谢组学研究策略,分别对患者与健康对照者的血清和尿液氨基酸水平进行分析,采用主成分分析与正交偏最小二乘-辨别分析(OPLS-DA)的多元统计分析方法进行建模,选择OPLS-DA模型的变量重要性投影值>1与 t检验的 P<0.05来筛选差异氨基酸代谢物,通过受试者工作特征(ROC)曲线分析和二元Logistic回归分析推断潜在标记物的诊断效能。 结果:研究筛选出丝氨酸、谷氨酸、天冬氨酸、异亮氨酸和甘氨酸等5种氨基酸代谢物在尿石症组与对照组的差异具有统计学意义( P<0.05),且与7条代谢通路相关联。将血清丝氨酸、谷氨酸、天冬氨酸、异亮氨酸和尿液甘氨酸、天冬氨酸组合成联合标志物组,其ROC曲线下面积为0.890,敏感度为78.0%,特异度为96.4%。 结论:共发现血清和尿液中的5种氨基酸可作为尿石症早期筛查诊断的生物标志物,为尿石症的分子基础研究提供一定的实验依据。

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abstracts:

Objective:To analyze the serum and urinary amino acid (AA) profiles of urolithiasis patients to explore the potential biomarkers for clinical screening and early diagnosis.Methods:Case-control study. Serum and urine samples were collected from 74 urolithiasis patients (aged 20-82 years, 41 men, 33 female) in the department of urology of the First Affiliated Hospital of Fujian Medical University and 35 healthy controls (HC, aged 22-80 years old, 20 men, 15 female) from the health examination center from February 2015 to October 2017. Serum and urinary AA levels of patients and HC were analyzed using Gas Chromatography-Mass Spectrometry (GC-MS) based metabolomic strategy. The multivariate statistical analysis methods of principal component analysis (PCA) and orthogonal partial least squares discrimination analysis (OPLS-DA) were employed for modeling. The variable importance projection (VIP) value of OPLS-DA model>1 and P<0.05 of t test were selected to screen the differential amino acid metabolites. The diagnostic capabilities of potential markers were evaluated by receiver operating characteristic (ROC) curve and binary logistic regression analysis. Results:Five AA metabolites including serine, glutamate, aspartic acid, isoleucine and glycine were found, which had statistically significant differences between the patient group and the control group ( P<0.05) and were associated with seven metabolic pathways. Serum serine, glutamate, aspartic acid, isoleucine and urine glycine and aspartic acid were combined into an integrated marker panel whose AUC value was 0.890, the sensitivity was 78.0%, and the specificity was 96.4%. Conclusion:Five amino acids in serum and urine could be used as an integrated biomarker panel for the clinical screening and early diagnosis of urolithiasis, which could provide some experimental basis for molecular urolithiasis research.

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作者: 高瑶 [1] 林芯馨 [1] 龚思思 [2] 张天闻 [3] 唐敏洁 [4] 张蓓英 [1] 陈敏 [1] 欧启水 [4] 毛厚平 [5]
期刊: 《中华检验医学杂志》2022年45卷5期 463-471页 ISTICPKUCSCD
栏目名称: 论著
DOI: 10.3760/cma.j.cn114452-20210720-00444
发布时间: 2022-07-31
基金项目:
国家自然科学基金 福建省自然科学基金 福建省大学生创新创业训练计划 National Natural Science Foundation of China Fujian Natural Science Foundation College Students' Innovation and Entrepreneurship Training Program of Fujian Province
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