基金项目:上海市科委资助项目(024119021)作者单位:200092上海第二医科大学附属新华医院,上海市儿科医学研究所内分泌、遗传代谢病科(顾学范、韩连书、高晓岚、叶军、邱文娟);北京大学第一医院儿科(杨艳玲)・论著・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顾学范 韩连书 高晓岚 杨艳玲 叶军 邱文娟 【摘要】 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104例,男77例,女27例,年龄(418±412)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104例标本中,检出阳性标本10例(916%),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,32羟基32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。
【关键词】 光谱分析,质量; 代谢缺陷,先天性; 氨基酸类; 肉碱A pilot study of selective screening for high risk children with inborn error of metabolism using tandem m ass spectrometry in China G U Xue2fan,H AN Lian2shu,G AO Xiao2lan,Y AN Yan2ling,YE Jun,QIU Wen2juan.Department o f Pediatric Endocrinology and G enetic Metabolic Diseases,Xinhua Hospital,Shanghai Second Medical Univer sity,Shanghai Institute for Pediatric Research,Shanghai200092,China 【Abstract】 Objective The new technology of tandem mass spectrometry is exerting a significant im pact on the diagnostics of inborn metabolic errors,and allows to detect a number of these dis orders in a single step.The aim of the present study was to establish a dry blood filter paper method for amino acid and acylcarnitine profiles test using tandem mass spectrometry and to apply the method for selective screening in high risk children with inborn error of metabolism.Method The study group consisted of104high risk cases of inborn error of metabolism from 5pediatric hospitals in Shanghai and Beijing were tested from N ovember2002to June2003;77were males and27 females,the means age was418±412years.These patients had mental retardation,slow development, psychological abnormalities,muscle hypotonia,jaundice,hepatosplenomegaly,recurrent v omiting,and boratory examinations suggested metabolic acidosis,hypoglycemia,hyperamm onemia and hyperlactacidemia.Phenylketonuria was excluded in this study by routine phenylalanine screening.The control group consisted of308children,170males and138females.The blood was collected on filter paper,punched and extracted into methanol s olution with stable is otope labeled internal standards,then derivatized with butanolic2HCl.A fter preparation,the sam ples were analyzed by tandem mass spectrometry(API2000).R esults T en of104 patients(916%)were positive in our selective screening program,including one with tyrosinemia,one with hom ocystinuria,one with hyperornithinemia,tw o with methylmalonic acidemia,one with propionic acidemia,one with32hydroxy232methylglutaryl2C oA lyase deficiency,tw o with medium chain acyl2C oA dehydrogenase deficiency, and one with carnitine palmitoyl trans ferase typeⅡdeficiency.Conclusion The authors established a fast, accurate and sensitive tandem mass spectrometry method for amino acid and acylcarnitine profiles analysis,nearly 30metabolic diseases including amino acid dis orders,organic acid dis orders and fatty acid oxidation dis orders could be detected,m ost of the diseases that cause death and disability represent preventable entities by early diagnosis and treatment.The results indicated the im portance of selective screening for high risk patients with inborn error of metabolism.【K ey w ords】 S pectrum analysis,mass; Metabolism,inborn errors; Amino acids; Carnitine 遗传性代谢病较多依赖生化与分子生物学方法,对技术和设备要求较高,往往是临床的诊治难点[1,2]。
