串联质谱技术在有机酸血症筛查中的应用研究

目的探讨串联质谱技术在遗传性代谢病高危儿童有机酸血症筛查和诊断中的意义。方法遗传性代谢病高危儿童1000例,男642例,女358例,年龄中位数为2岁(5天～15岁),手指采血,滴于滤纸片上。取直径3mm的干血滤纸片放入96孔聚丙烯板,经含酰基肉碱内标的甲醇萃取,盐酸正丁醇衍生后,进行串联质谱分析。结果1000例高危儿童中筛查并诊断有机酸血症40例(4．0％),其中甲基丙二酸血症20例,丙酸血症6例,异戊酸血症3例,戊二酸血症Ⅰ型3例,戊二酸血症Ⅱ型3例,β-甲基巴豆酰辅酶A羧化酶缺乏症1例,β-羟基．3．甲基戊二酰辅酶A裂解酶缺乏症1例,生物素酶缺乏症2例,β-酮硫解酶缺乏症1例。患儿临床表现主要为运动和智力发育落后、抽搐、意识障碍、肌张力低下、呕吐和喂养困难等。常规实验室检查主要表现为酸中毒、高血氨、高乳酸血症和低血糖等。结论串联质谱技术能够通过检测血滤纸片中的不同酰基肉碱浓度,对部分有机酸血症进行快速筛查和诊断,结合尿气相色谱／质谱技术,有助于提高有机酸血症诊断的准确性。     

气相-色谱质谱法尿有机酸检测诊断遗传代谢病患儿的疾病谱

目的了解气相色谱-质谱技术检测尿有机酸水平诊断的遗传代谢病患儿的氨基酸、有机酸、脂肪酸氧化代谢病疾病谱。方法回顾性分析2005年1月至2021年12月上海交通大学医学院附属新华医院诊断为遗传代谢病的2 461例患儿的疾病谱, 患儿均通过气相色谱-质谱尿有机酸谱检测, 并结合血串联质谱氨基酸及酰基肉碱检测结果及基因变异检测结果诊断。结果 2 461例患儿中男1 446例, 女1 051例, 共有32种遗传代谢病, 其中氨基酸代谢病10种662例(26.9%), 常见的前6种疾病为高苯丙氨酸血症、希特林蛋白缺乏症、鸟氨酸氨甲酰转移酶缺乏症、枫糖尿病、尿黑酸尿症及酪氨酸血症-Ⅰ型;有机酸血症17种1 683例(68.4%), 常见的前6种疾病为甲基丙二酸血症、丙酸血症、戊二酸血症-Ⅰ型、异戊酸血症、3-甲基巴豆酰辅酶A羧化酶缺乏症及多种羧化酶缺乏症;脂肪酸β氧化代谢病5种116例(4.7%), 常见的前2种疾病为多种酰基辅酶A脱氢酶缺乏症和短链酰基辅酶A脱氢酶缺乏症。结论气相色谱-质谱技术尿有机酸谱检测诊断的疾病中有机酸血症最常见, 然后为氨基酸代谢病及脂肪酸氧化代谢病。     

尿素酶预处理-气相色谱-质谱法选择性筛查遗传代谢病高危患儿327例初步研究

目的　通过对遗传代谢病高危患儿尿液成分进行生化分析 ,筛查遗传代谢病 ,为临床诊断和治疗提供实验依据。方法　收集遗传代谢病高危患儿尿液标本 ,经去尿素、加内标、除蛋白、真空干燥、三甲基硅烷基衍生等处理后 ,应用气相色谱 -质谱联用仪分析尿液中有机酸、氨基酸、糖类、多醇、嘌呤、嘧啶等成分。这一流程在国际上被称为尿素酶预处理 气相色谱 质谱法。结果　对来自中国大陆 6省、区和直辖市的 3 2 7例遗传代谢病高危患儿的尿液标本进行检测 ,共筛查出遗传代谢病 16种 2 7例 ,阳性率为 8.2 6% ,其中高苯丙氨酸血症、甘油尿症和Leigh综合征各 3例 ,丙酸血症、甲基丙二酸尿症、vonGierke病、果糖 1,6 二磷酸酶缺陷病、果糖尿症各 2例 ,多种羧化酶缺陷病、戊二酸血症Ⅰ型、枫糖尿病、高甘氨酸血症、3 氨基异丁酸尿症、半乳糖血症、瓜氨酸血症Ⅱ型及Fanconi综合征各 1例。经临床干预虽然仍有部分患儿预后不良 ,但多种羧化酶缺陷病、甲基丙二酸尿症、半乳糖血症等患儿获得较好的治疗效果。其余患儿的病情有待追踪观察。结论　应用尿素酶预处理 气相色谱 质谱法分析尿液成分 ,是筛查某些遗传代谢病的有效方法 ,检测结果可为患儿的诊断和治疗提供有效指导。     

串联质谱技术在有机酸血症鉴别诊断中的应用

目的探讨串联质谱技术在有机酸血症诊断和鉴别诊断中的作用。方法对2566例遗传性代谢病高危儿童的干血滤纸片进行串联质谱分析,结合尿气相色谱质谱有机酸分析、酶活性测定和基因突变分析,诊断有机酸血症。比较不同有机酸血症间的串联质谱结果差异。结果共确诊有机酸血症82例(3.2%),其中甲基丙二酸血症(MMA)44例,丙酸血症(PA)10例,生物素酶缺乏症4例和全羧化酶合成酶缺乏症2例;患儿丙酰肉碱(propinoylcar-nitine,C3)水平均高于健康儿童上限值(4.0μmol/L)。PA和MMA患儿C3与游离肉碱(C0)比值(C3/C0)、C3与乙酰肉碱(C2)比值(C3/C2)也高于健康儿童上限值;PA患儿同时伴有甘氨酸增高,甲基丙二酸血症患儿甘氨酸正常;生物素酶缺乏症和全羧化酶合成酶症患儿伴3"羟基"异戊酰肉碱(3"hydyoxyl"isovalerylcarnitine,C5"OH)水平增高;3"甲基巴豆酰辅酶A羧化酶缺乏症(5例)、3"羟基"3"甲基戊二酰辅酶A裂解酶缺乏症(3例)和β"酮硫解酶缺乏症(4例)患儿C5"OH水平也增高,前两种疾病仅表现为C5"OH增高,后一种疾病同时伴有异戊烯酰基肉碱水平增高;异戊酸血症5例,仅表现为异戊酰肉碱(C5)增高;戊二酸血症(GA)"Ⅰ型(3例)仅表现为戊二酰肉碱(C5DC)增高,GA"Ⅱ型(2例)除C5DC增高外,伴有多种酰基肉碱增高,两者可作鉴别。结论通过串联质谱酰基肉碱、酰基肉碱比值和氨基酸分析,可对部分有机酸血症进行诊断和鉴别诊断;部分有机酸血症需结合尿气相色谱质谱有机酸分析进行诊断。     

先天性遗传代谢病的早期诊断

目的提高儿科医生对新生儿期遗传代谢病的认识,做到早期诊断、早期治疗.方法自2003年9月至2004年9月,根据临床表现确定18名遗传代谢病高危患儿,用"滤纸片代"将采集的尿标本外寄进行气相色谱-质谱(GC/MS)分析,筛查遗传代谢病.结果 18例高危儿中确诊为遗传代谢病5例,分别为戊二酸尿症Ⅱ型1例(46h,男),鸟氨酸氨甲酰转移酶缺陷1例(66h,男),枫糖尿病1例(8 d,男),甲基丙二酸血症1例(13 d,男),丙酸血症1例(21 d,女),并对其临床特点进行归纳总结.结论掌握新生儿遗传代谢病临床特点,对高危儿早期进行尿GC/MS分析,可以早期诊断遗传代谢病,有利于优生优育.     

串联质谱技术在遗传性代谢病高危儿童筛查中的初步应用

目的　开展遗传性代谢病高危儿童筛查检测。方法　临床疑诊遗传性代谢病儿童 10 4例 ,男 77例 ,女 2 7例 ,年龄 (4 8± 4 2 )岁。手指采血 ,滴于专用采血滤纸上。干血滤纸片与含已知量的氨基酸、酰基肉碱同位素内标一起 ,经甲醇萃取 ,盐酸正丁醇衍生后 ,用串联质谱仪分析血片中氨基酸谱和酰基肉碱谱及其浓度。结果　在 10 4例标本中 ,检出阳性标本 10例 (9 6 % ) ,其中酪氨酸血症1例 ,同型胱氨酸血症 1例 ,高鸟氨酸血症 1例 ,甲基丙二酸血症 2例 ,丙酸血症 1例 ,3 羟基 3 甲基戊二酸裂解酶缺乏症 1例 ,中链酰基辅酶A脱氢酶缺乏症 2例 ,酰基肉碱转移酶Ⅱ缺乏症 1例。结论串联质谱技术可检测 30余种遗传性代谢病 ,包括氨基酸代谢紊乱、有机酸代谢紊乱和脂肪酸代谢紊乱性疾病 ,高危遗传性代谢病患儿的筛查有重要价值     

先天畸形及代谢缺陷

040447尿滤纸片法气相色谱一质谱分析技术在遗传性代谢病高危筛查诊断中的应用/罗小平…//中华儿科杂志一2003,41(4)一245一248 在352例标本中共检出遗传性代谢病34例,包括甲基丙二酸血症,Q一酮戊二酸尿症、酪氨酸血症I型及二梭酸尿症等。常见临床症状和体征为:智能障碍、惊厥、肌张力异常和黄疽;常规实验室检查多见代谢性酸中毒、低血糖、高血氨等。参16(何燕) 040448新生儿出生缺陷76例临床分析/解雪梅…//陕西医学杂志一2003 .32(7)一596~597 3年间共出生新生儿5876人,其中有出生缺陷76例,占总出生人数的12.9%。。76例新生儿缺陷中房/室…     

先天性遗传代谢病的早期诊断

目的提高儿科医生对新生儿期遗传代谢病的认识，做到早期诊断、早期治疗。方法自2003年9月至2004年9月，根据临床表现确定18名遗传代谢病高危患儿，用“滤纸片代”将采集的尿标本外寄进行气相色谱．质谱(GC／MS)分析，筛查遗传代谢病。结果18例高危儿中确诊为遗传代谢病5例，分别为戊二酸尿症Ⅱ型1例(46h，男)，鸟氨酸氨甲酰转移酶缺陷1例(66h，男)，枫糖尿病1例(8d，男)，甲基丙二酸血症1例(13d，男)，丙酸血症1例(21d，女)，并对其临床特点进行归纳总结。结论掌握新生儿遗传代谢病临床特点，对高危儿早期进行尿GC／MS分析，可以早期诊断遗传代谢病，有利于优生优育。     

新生儿期氨基酸、有机酸及脂肪酸氧化代谢病疾病谱分析

目的了解中国新生儿期发病的氨基酸、有机酸和脂肪酸氧化代谢病疾病谱,以提高对此类疾病的认识。方法回顾性分析2003年2月—2012年5月就诊的2 547例疑似遗传性代谢病新生儿的血串联质谱和尿气相色谱检测结果,并对其中确诊的127例新生儿的疾病谱及临床表现进行分析。结果在2 547例新生儿中诊断氨基酸、有机酸和脂肪酸氧化代谢病127例(5.0%)。其中氨基酸代谢病37例(29.1%),以鸟氨酸氨甲酰基转移酶缺乏症(16例)和枫糖尿病(14例)多见;有机酸代谢病76例(59.8%),以甲基丙二酸血症(50例)最常见,其次为丙酸血症(13例)和异戊酸血症(10例);脂肪酸氧化代谢病14例(11.0%),以多种酰基辅酶A脱氢酶缺乏症多见(6例)。结论新生儿期甲基丙二酸血症最常见,其次为鸟氨酸氨甲酰基转移酶缺乏症、枫糖尿病、丙酸血症和异戊酸血症。儿科医师尤其是新生儿科医师应重视利用串联质谱和气相质谱技术进行遗传代谢病检测,以达到早诊断、早治疗的目的。     

气相色谱-质谱法对高危婴幼儿遗传性代谢疾病筛查的研究

目的：研究气相色谱－质谱法对高危婴幼儿遗传性代谢疾病筛查的意义的价值,并对阳性患儿的临床特点进行分析。方法：对705名原因不明的智力－运动发育迟缓、倒退及疑有遗传代谢病的婴幼儿,采用气相色谱－质谱（GC－MS）方法进行尿液化学分析。结果：705例中尿化学分析异常者65例（9．2％）,均为遗传性代谢疾病患儿,其中半乳糖血症11例,苯丙酮尿症、甲基丙二酸尿症各10例,范可尼综合征3例、戍二酸、赖氨酸尿症、疑似同型丝氨酸尿症各2例,果糖1．6二磷酸酶缺乏症、焦谷氨酸尿症、长链脂肪酸尿症及神经母细胞瘤、鸟氨酸尿症、多羧酸酶缺乏症各1例,单纯性糖尿2例,糖尿病2例,乳酸及酮尿症15例。对筛查出的数种疾病患儿进行合理的治疗均取得明显效果。结论：（1）对不明原因的智力－运动发育及倒退的婴幼儿应进行遗传性代谢疾病的筛查。（2）GC－MS检测技术方法准确,敏感性高,特异性强,是筛查遗传性代谢疾病的有效手段。（3）婴幼儿遗传性代谢疾病的筛查应尽早进行,扩大新生儿遗传性代谢疾病筛查的覆盖面,增加筛查病种是非常重要的。     

Biochemical diagnosis of rare hereditary metabolic disorders. Experiences from a patient sample of the Innsbruck University Pediatric Clinic 1984-1987

The diagnosis of inherited metabolic diseases requires an intensive cooperation between the clinician, the clinical chemist and the biochemist to select patients for an adequate screening program, to avoid unnecessary expensive investigations and to achieve a final molecular diagnosis. Often a cooperation with specialized laboratories even in different countries is necessary. 29 patients with an inherited metabolic disorder have been evaluated at the Children's Hospital, University of Innsbruck, between 1984 and 1987. Seven patients with a disturbance in carbohydrate metabolism incl. pyruvate oxidation, nine with a defect in amino or organic acid metabolism, eleven patients with a lysosomal storage disorder, one patient with steroid-sulfatase deficiency and one with M. Wilson have been found. In nine of these 29 patients an organic aciduria was detected, four of them had lactic aciduria due to a mitochondrial defect of the oxidative energy metabolism. 21 children corresponded to a well-known classic phenotype, eight represented very rare or unique, not previously described clinical forms of a disease. In one third of the cases a cooperation with a foreign institute was necessary. The presentation of the most important biochemical investigations in children with inherited metabolic defects should give a survey of the diagnosis of such patients at a pediatric centre and should furthermore stimulate the development of a common diagnostic procedure program and the cooperation and coordination of the laboratories involved in the diagnosis of inherited metabolic disorders in Austria.     

Differential diagnosis of the symptom of lactic acidosis in childhood

Transitory Lactic acidemia may be found under very different conditions, eg. hyperventilation, hypoxia, strenuous muscular work, diabetic ketoacidosis, malignancies and totally parenteral alimentation. In contrast chronic, permanent lactic acidemia and lactic aciduria may point to one of several and usually severe inherited metabolic diseases of gluconeogenesis, pyruvate oxidation, the Krebs cycle or the respiratory chain. Clinical hints for a stepwise differential diagnosis are presented.     

有机酸血（尿）症及其临床处理

有机酸血（尿）症是临床最常见的一类遗传代谢病,目前已经发现约50余种,多数在新生儿期或婴幼儿期发病.临床上多表现为顽固性代谢性酸中毒、发作性呕吐、喂养困难、肌张力低下、惊厥和意识障碍等.由于本类疾病临床没有特异性,若不能早期诊断和治疗,易出现猝死或不可逆转的神经系统损伤.利用气相色谱-质谱联用技术和（或）串联质谱技术对疑似有机酸血（尿）症患儿进行早期生化诊断是改善患儿预后和挽救患儿生命的关键.     

15851例遗传代谢病高危患儿的临床分析

目的探讨尿气相色谱-质谱法在遗传代谢病(IMD)高危患儿筛查中的应用价值,以及确诊IMD的疾病谱和确诊IMD患儿的临床特点,为临床诊断和治疗提供参考。方法回顾性分析2012年2月至2016年12月在天津市儿童医院就诊的15 851例IMD高危患儿且接受了尿气相色谱-质谱法检测的患儿的临床资料。结果 15 851例IMD高危患儿中,检出代谢异常5 793例,占36.55%。确诊为IMD 117例(0.74%),其中甲基丙二酸血症77例(65.8%)。新生儿期IMD确诊患儿的临床表现主要为黄疸、代谢性酸中毒、肌张力异常、喂养困难、反应差、嗜睡或昏迷等,非新生儿期IMD确诊患儿的临床表现主要为精神运动发育落后、代谢性酸中毒、抽搐、反复呕吐、贫血等。结论气相色谱-质谱法是对IMD高危患儿筛查的有效方法;甲基丙二酸血症是最常见的IMD; 新生儿期和非新生儿期IMD确诊患儿的临床表现有不同。     

Biotinidase deficiency: a cause of subacute necrotizing encephalomyelopathy (Leigh syndrome). Report of a case with lethal outcome

An unusual clinical course of a patient with biotinidase deficiency, causing Leigh syndrome, is reported. Laryngeal stridor was the major presenting symptom followed by progressive neurologic deterioration and death at the age of 21.5 mo. Absence of skin and hair abnormalities as well as of organic aciduria delayed the correct diagnosis. Necropsy revealed subacute necrotizing encephalopathy (Leigh syndrome). Carboxylase activities (propionyl CoA carboxylase, 3-methylcrotonyl-CoA carboxylase, pyruvate carboxylase) measured in lymphocytes 1 day before death were decreased to 10% of normal values. Propionyl-CoA carboxylase was shown to be the only stable carboxylase in human postmortem tissue; in our patient it was moderately decreased in postmortem liver (29% of control) and kidney (42%), but severely decreased in brain (3%). These findings might explain the severity of neurological symptoms in the absence of marked organic aciduria. They indicate that in biotinidase deficiency the CNS may become biotin depleted earlier and more severely than other organs. Biotinidase deficiency should be included in the differential diagnosis of Leigh syndrome and of unexplained respiratory problems.     

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??Methylmalonic acidemia??also known as methylmalonic aciduria??is the most common disease of organic acids metabolic disorders. The patients usually present with multi-organic damage. Neurological diseases are common findings. In some patients??kidney disease is one of the complication or the only symptom. Kidney damage due to methylmalonic acidemia needs much attention. Renal damage caused by methylmalonic acidemia is mainly manifested as tubulointerstitial injury. The common manifestations are hematuria??proteinuria??edema and hypertension. If left untreated??the pregressive disease will lead to renal failure. The detection of plasma total homocysteine??blood amino acids??acyl-carnitines and urinary organic acids and genetic analysis are the important methods of etiological diagnosis with methylmalonic acidemia. Most of the patients of methylmalonic acidemia are cobalamin-effective. The renal diseases are reversible. Many patients are improved or get recovery. For the patients with unexplained renal diseases??the differential diagnosis should be paid attention to. Early diagnosis and early intervention are keys to improving the outcome of the patients with methylmalonic acidemia.     

Seven years of experience with selective screening for organic acidurias

Between 1975 and 1981 nearly 9000 patients with suspected inherited metabolic diseases were investigated by a selective screening procedure including, apart from simple tests for ketone bodies, sugars and SH-containing compounds, high voltage electrophoresis of amino acids as well as gas liquid chromatography and gas liquid chromatography-mass spectrometry of the organic acids. Fifty-two cases with 18 different inborn errors of metabolism were detected. The effectivity index was calculated to be 0.6% or 1 case in about 170 requests. From the presented and from already existing data in the literature the overall incidences for all organic acidurias together and for propionic acidemia separately were appraized to be 1:10000 and 1:50000, respectively. About half of the patients diagnosed by this screening may benefit from the diagnosis.     

维生素B2治疗有效的晚发型戊二酸尿症Ⅱ型

目的 探讨戊二酸尿症Ⅱ型(glutaric aciduria typeⅡ,GAⅡ)的诊断和治疗。方法 应用气相色谱-质谱分析技术(GC-MS)对2例肌无力患儿的尿滤纸片标本进行有机酸分析。确诊为GAⅡ型后,在低蛋白、低脂肪、高碳水化合物饮食控制的基础上,应用VitB2进行治疗,并观察治疗前后临床、生化及尿有机酸分析等方面的变化。结果 (1)两患儿尿滤纸片标本有机酸分析检出大量戊二酸、异戊酰甘氨酸、乙基丙二酸及己二酸、辛二酸、癸二酸等二羧酸,提示为戊二酸尿症Ⅱ型;(2)经饮食控制和VitB2治疗后,2例患儿的肌无力症状明显好转,肌力明显增强,四肢肌张力恢复正常,尿有机酸分析显示上述有机酸的排出量逐步降低、甚至消失,血清谷-草转氨酶、肌酸激酶以及乳酸脱氢酶水平均较治疗前显著降低。血氨基本降至正常水平。结论 戊二酸尿症是一种严重的先天代谢障碍性疾病。部分本症患儿在低蛋白、低脂肪、高碳水化合物饮食控制的基础上,应用VitB2进行治疗,可取得较好的临床效果。     

Metabolic disorders of branched-chain amino acids: most common forms of organic aciduria in the neonatal period

Defects in the branched chain amino acid metabolism are the most common forms of organic aciduria. Two thirds of the cases manifest themselves during the neonatal period, most of them with an acute onset. Prompt diagnosis of organic acidurias is a task of the pediatrician and the neonatologist and depends on their early identification of children with suspect clinical symptoms. Between 1984 and 1987 9 patients presented with an organic aciduria at the Pediatric Department of the University of Innsbruck, 7 of them were neonates. 4 of these 7 children had a defect in the branched chain amino acid metabolism, 3 with propionic acidemia, one with maple syrup urine disease. The remaining 3 children presented with lactic aciduria. In all our patients diagnosis was performed by combined gas chromatography and mass spectrometry of spontaneous urine samples. Diagnostic procedures and therapeutic measures applied in the acute metabolic crisis are presented. Continuous arteriovenous hemofiltration has been found to be an efficient method for eliminating toxic metabolites in intractable metabolic acidosis.