成纤维细胞体外培养结合串联质谱酰基肉碱谱诊断极长链酰基辅酶A脱氢酶缺陷症  

Diagnosis of Very-Long-Chain Acyl-CoA Dehydrogenase Deficiency Using Tandem Mass Spectrometry Combined with in-Vitro Acylcarnitine Profiling Assay

作　　者：李红[1,2] 马立燕[1] 马金海[1] 福田诚司 长谷川有纪 小林弘典 山口清次 

机构地区：[1]宁夏医科大学附属医院儿科,银川750004 [2]日本岛根大学医学部小儿科,日本岛根6938501

出　　处：《实用儿科临床杂志》2009年第20期1560-1562,共3页Journal of Applied Clinical Pediatrics

基　　金：Program for the Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation(NIBIO)of Japan

摘　　要：目的研究成纤维细胞体外培养结合串联质谱酰基肉碱谱分析对极长链酰基辅酶A脱氢酶(VLCAD)缺陷症的诊断价值。方法取健康对照标本6例,VLCAD缺陷症患儿标本9例(包括临床严重型2例,中间型3例及肌病型4例。男5例,女4例)。以棕榈酸(C16酸)为培养溶媒底物在37℃培养箱中培养成纤维细胞株96h,应用串联质谱分析相应培养液中的酰基肉碱谱。结果与健康对照组比较,VLCAD缺陷症患儿乙酰肉碱(C2)明显降低,相应长链酰基肉碱异常增高,其中肉豆寇烯酰肉碱(C14)-酰基肉碱在临床严重型患儿中增高明显,月桂酰肉碱(C12)-酰基肉碱在临床轻型病例(包括中间型和肌病型)中增高更显著。长链酰基肉碱之和(ΣLC)及其与乙酰肉碱之比值(ΣLC/C2),VLCAD缺陷症患儿均明显高于健康对照。结论成纤维细胞体外特定条件培养结合串联质谱酰基肉碱谱分析,可诊断和鉴别VLCAD缺陷症及其临床分型,C12-酰基肉碱和C14-酰基肉碱可作为诊断标记。Objective To study the application of tandem mass spectrometry （MS/MS） combined with in -vitro acylcarnitine profiling （AP） assay in diagnosing very - long - chain acyl - CoA dehydrogenase （VLCAD） deficiency. Methods Human fibroblasts from normal controls ＂rod VLCAD deficiency children （male 5 cases, female 4 cases） including 2 cases of severe, 3 cases of intermediate and 4 cases of myopathic clinical symptoms,were cultured in medium loaded with unlabelled palmitic acid for 96 h at 37 ℃. The quantitative AP in the medium wasanalyzed by using tandem MS. Results Compared with normal control, acetylcarnitine （ C2 ） decreased significantly and long - chain acylcarnitines,especially tetradecanoglcarnitine（C14） -acylcarnitine in clinical severe symptoms and dodecanoylcarnitine（C12） - acylcarnitine in mild symptoms （including intermediate and myopathic symptoms）, elevated significantly in VLCAD deficiency. Indeed, the sum of long - chain acylcarnitine （∑LC） and the ratio of ∑LC/C2 increased significantly in VLCAD deficiency. Conclusions Tandem MS - based quantitative AP derived from medium, which was loaded with unlabelled palmitic acid for culturing fibroblasts, is a useful and simple method for distinguishing VLCAD deficiency and its clinical symptoms,and C12 - and C14- acylcarnitine can be diagnostic markers.

关 键 词：极长链酰基辅酶A脱氢酶缺陷症 串联质谱 酰基肉碱 成纤维细胞 

分 类 号：R725.8[医药卫生—儿科]