细胞学结合串联质谱诊断多种酰基辅酶A脱氢酶缺乏症的价值

目的 研究应用串联质谱技术分析细胞培养液中不同链长酰基肉碱谱的变化对多种酰基辅酶A脱氢酶缺乏症（multiple acy-CoA dehydrogenase deficiency,MADD）的诊断价值.方法 取健康对照标本20例,MADD患儿皮肤成纤维细胞标本9例（包括Ⅰ型3例,Ⅱ型3例和Ⅲ型3例.在37℃培养条件下,用特殊培养底物即加入棕榈酸（C16酸）的培养液中培养成纤维细胞96 h,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,MADD患儿乙酰肉碱（C2）明显降低,而所有病人月桂酰肉碱（C12）、肉豆蔻酰肉碱（C14）、及棕榈酰肉碱（C16）等长链酰基肉碱均异常增高,与健康对照组相比中链酰基肉碱C6、C8、C10在重型（Ⅰ型）病例异常降低,而Ⅱ型与Ⅲ型中链酰基肉碱C6、C8、C10与健康对照比明显升高;其中棕榈酸酰基肉碱（C16）在Ⅰ型患儿中明显增高,肉豆寇烯酰肉碱（C14）及月桂酰肉碱（C12）在临床Ⅲ型和Ⅱ型病例增高更显著.结论 应用串联质谱酰基肉碱谱分析结合特定培养液培养皮肤成纤维细胞可用于MADD的诊断和临床分型.

Diagnosis and Classifying of Multiple Acy-CoA Dehydrogenase Deficiency Using Tandem Mass Spectrometry Combined with In-vitro Acylcarnitine Profiling Assay

Objective To explore the application of tandem mass spectrometry （MS/MS） combined with in-vitro acylcarnitine profiling （AP） assay in diagnosing.multiple acy-CoA dehydrogenase deficiency Methods Human fibroblasts from 20 normal controlls and 9 multiple acy-CoA dehydrogenase deficiency children including 3 cases of,3 cases of and 3 cases of symptoms,were cultured in medium loaded with unlabelled palmitic acid for 96h at 37℃.The quantitative AP in the medium was analyzed by using tandem MS.Results Compared with normal controll,acetylcarnitine （C2） decreased significantly in patients while longchain acylcarnitines,especially palmitic carnitine （C16）,dodecanoylcarnitine （C12） and tetradecanogl carnitine （C14） elevated significantly in patients with multiple acy-CoA dehydrogenase deficiency.Conclusion Tandem mass spectrometry combined with culturing human being skin fibroblasts can be used to diagnosis and classify multiple acy-CoA dehydrogenase deficiency.