尿滤纸片法气相色谱-质谱分析技术在遗传性代谢病高危筛查诊断中的应用  

Application of gas chromatography-mass spectrometry analysis on urine filter paper in the high-risk screening and diagnosis of inherited metabolic diseases

作　　者：罗小平[1] 王慕逖[1] 魏虹[1] 梁雁[1] 王宏伟[1] 林汉华[1] 董永绥[1] 刘皖君[1] 方俊敏[1] 宁琴[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院儿科

出　　处：《中华儿科杂志》2003年第4期245-248,T001,共5页Chinese Journal of Pediatrics

基　　金：国家自然科学基金委员会国家杰出青年科学基金( 30 12 50 19);卫生部临床学科重点建设项目( 970 70 2 4 0)

摘　　要：目的　建立一种适合我国国情的有机酸尿症及其他遗传性代谢病的高危筛查诊断方法。方法　应用滤纸片收集尿标本 ,建立尿有机酸及氨基酸代谢产物的萃取、衍化和气相色谱 质谱分析方法 ;成立遗传代谢性疾病高危筛查诊断协作网络并开展工作。结果　 352例样本中共检出阳性病例 34例 (9 66 % ) ,病种包括甲基丙二酸血症 6例 ,α 酮戊二酸尿症 5例 ,酪氨酸血症Ⅰ型 4例 ,二羧酸尿症 4例 ,多种羧化酶缺乏症 3例 ,苯丙酮尿症 3例 ,高乳酸血症 3例 ,丙酸血症 2例 ,鸟氨酸氨甲酰基转移酶缺乏症 1例 ,乙基丙二酸 己二酸尿症 1例 ,戊二酸尿症Ⅱ型 1例和 3 甲基巴豆酰羧化酶缺乏症 1例。常见的临床症状和体征包括智能发育落后、惊厥、肌张力异常和黄疸。常规实验室检查多见代谢性酸中毒、低血糖和高血氨等异常。结论　尿有机酸气相色谱 质谱分析为遗传性代谢病高危筛查诊断的重要技术。应用滤纸片收集尿标本后在遗传代谢病诊断中心进行检测为实用而有效的方法。Objective To establish a specific procedure for the high risk screening and diagnosis of organic acidurias and other inherited metabolic diseases in China Methods A nation wide network for the high risk screening and diagnosis of genetic metabolic diseases was formed to facilitate the collaboration Urine samples were collected using filter paper from patients with clinical symptoms suspicious of inherited metabolic diseases The samples were eluted with distilled water and internal standards were added Samples were treated with hydroxylamine hydrochloride to form oximes to improve the recoveries of 2 ketoacids Urinary organic acids were extracted with ethyl acetate and diethyl ether under acidic condition After dehydration, the combined organic phase was evaporated to dryness with nitrogen The residues were added with BSTFA+1%TMCS and heat incubated to form the trimethylsilyl derivatives, and then were analyzed on an Agilent 5 890/5 973N gas chromatography mass spectrometer (GC MS), with a 7 683 series auto sampler The peaks were identified by reference to a mass spectral library Results Totally 352 samples were collected from the network collaborating hospitals since 2001 Thirty four (9 66%) cases of various inherited metabolic diseases were diagnosed with an age range of 2 days to 14 years The disease profile was consisted of methylmalonic acidemias (6), α keto glutaric aciduria (5), tyrosinemia type Ⅰ(4), dicarboxylic aciduria (4), multiple carboxylase deficiency (3), phenylketonuria (3), lactic acidemia (3), propionic acidemia (2), ornithine transcarbamoylase deficiency (1), ethylmalonic adipic aciduria (1), glutaric aciduria type Ⅱ (1) and 3 methylcrotyl CoA carboxylase deficiency (1) The most common clinical symptoms and signs included mental and developmental retardation, convulsion,musculotonic abnormality and jaundice Routine laboratory tests often revealed metabolic acidosis, hypoglycemia and hyperammonemia, etc Conclusion Urine organic acids analysis by

关 键 词：尿滤纸片法 气相色谱-质谱分析技术 遗传性代谢病 诊断 应用 

分 类 号：R596[医药卫生—内科学] R58[医药卫生—临床医学]