羊水同型半胱氨酸检测与质谱检测联合产前诊断甲基丙二酸血症  

Prenatal diagnosis of methylmalonic acidemia using test of homocysteine and mass spectrometry in amniotic fluid

作　　者：陈婷[1] 韩连书[1] 季文君[1] 徐烽[1] 叶军[1] 邱文娟[1] 张惠文[1] 王瑜[1] 龚珠文[1] 顾学范[1] 

机构地区：[1]上海交通大学医学院附属新华医院、上海市儿科医学研究所内分泌/遗传科,200092

出　　处：《中华检验医学杂志》2017年第11期880-884,共5页Chinese Journal of Laboratory Medicine

基　　金：国家重点研发计划（2016YFC0901505）

摘　　要：目的 探讨羊水同型半胱氨酸检测联合液相色谱串联质谱（LC-MS/MS）及气相色谱质谱（GC/MS）检测对cblC型甲基丙二酸血症（MMA）产前诊断的价值.方法 回顾性研究.收集2014年1月至2017年3月于上海市儿科医学研究所进行产前诊断的187名孕妇检测结果,其中78例先证者为cblC型MMA患儿,109例为非有机酸血症孕妇（对照组）,于妊娠16~24周抽羊水,LC-MS/MS检测羊水丙酰肉碱（C3）和乙酰肉碱（C2）水平,GC/MS检测羊水甲基丙二酸、甲基枸橼酸水平,采用免疫荧光偏振法检测羊水同型半胱氨酸浓度,部分孕妇经培养羊水细胞进行MMACHC基因突变检测,本研究结果采用Mann-Whitney U秩和检验以及Kruskal-Wallis H检验进行相关统计学分析.结果 78例先证者为cblC型MMA的孕妇中,24例胎儿诊断为患儿（阳性组）,54例排除患病（阴性组）.阳性组HCY水平、C3水平、C3/C2值、甲基丙二酸水平及甲基枸橼酸水平均显著高于参考值、阴性组以及对照组（P值均为0.00）,差异有统计学意义;基因确诊胎儿为患儿的孕妇羊水代谢物检测均为阳性;孕妇羊水代谢物检测均为阴性者,基因检测结果均排除胎儿为患儿.另有2例胎儿基因检测结果为仅一个突变,而质谱和同型半胱氨酸结果均阳性.结论 羊水同型半胱氨酸检测联合LC-MS/MS、GC/MS检测可对cblC型MMA进行产前诊断,有助于弥补先证者基因突变不明的孕妇进行产前诊断.Objective To explore the value of the combination of homocysteine analysis, liquid chromatography tandem mass spectrometry（LC-MS/MS）and gas chromatography mass spectrometry（GC/MS）in the prenatal diagnosis of combined methylmalonic acidemia and homocystinuria（cblC defect）in amniotic fluid.Methods This is a retrospective study of 187 cases of pregnancies that came to our hospital for prenatal diagnosis between 2014/01-2017/03,among which 78 cases′probands were cblC defect patients and 109 cases′probands were not organic academia patients（control group）.Amniotic fluid samples from pregnant women were obtained at 16 -24 weeks of gestation.Propionylcarnitine（C3）and acetylcarnitine （C2）were measured by LC-MS/MS, methylmalonic acid and methylcitric acid were analyzed by GC /MS, and homocysteine was determined by fluorescence polarization immunoassay.Some pregnancies received MMACHC gene sequencing with cultured cells from amniotic fluid.Data were analyzed using Mann-Whitney U and Kruskal-Wallis H tests.Results Among those 78 pregnant women whose probands were diagnosed to be cblC defect,24 cases were diagnosed to be cblC defect（positive group）and 54 pregnant women were diagnosed to be negative（negative group）.In positive group, levels of homocysteine, C3, C3/C2, methylmalonic acid and methylcitric acid were all significantly higher than their normal reference ranges, negative group and control group（P values are 0.00）.Cases that were diagnosed to be cblC defect by MMACHC gene sequencing were all turned out to be positive in the tests of the above metabolites in amniotic fluid.Cases with negative results of the metabolites were all excluded to be cblC defect by gene sequencing. Besides,2 cases of pregnancies were diagnosed to be positive by homocysteine and mass spectrometric analysis while only one mutation were detected by gene sequencing.Conclusions The combination of homocysteine, LC-MS/MS and GC/MS analysis in amniotic fluid turns out to be reliable for prenatal diagnosis of c

关 键 词：氨基酸代谢障碍 先天性 产前诊断 羊水 高半胱氨酸 色谱法 液相 串联质谱法 气相色谱-质谱法 

分 类 号：R446.1[医药卫生—诊断学] R714.5[医药卫生—临床医学]