Citrin缺陷导致的新生儿肝内胆汁淤积症患儿热点突变基因筛查及生化改变研究

发布时间：2018-11-05 09:34

【摘要】：目的：了解深圳地区Citrin缺陷导致的新生儿肝内胆汁淤积症(NICCD)患儿常见热点突变基因特点以及纯合突变和杂合突变之间生化代谢改变关系,探讨NICCD患儿的临床表现及生化改变特征,为NICCD患儿的早期诊断治疗提供理论依据。方法：应用高分辨率熔解曲线(HRM)对临床可疑NICCD患儿进行四个热点突变基因类型(851del4,1638ins23, IVS6+5G＞A和IVS16ins3kb)进行筛查,并结合临床对基因学确诊患儿分别采用气相色谱质谱技术(GC/MS)和串联质谱技术(MS/MS)检测尿液及血液中的糖类、氨基酸及脂肪酸等代谢产物,同时检测肝代谢、肝酶、血氨、乳酸、甲胎蛋白、血脂等常规生化指标并分析。结果：360名可疑患儿中共筛查并确诊27例,阳性率为7.5%。四个热点突变中851del4突变占50.0%,1638ins23占13.0%,IVS16ins3kb占7.4%,IVS6+5GA占7.4%。纯合突变与杂合突变患儿之间体内的瓜氨酸,酪氨酸,苯丙氨酸,乙酰肉碱(C2),肉豆蔻酰肉碱(C14),棕榈酰肉碱(C16)浓度均无明显的差异性(P0.05)。27例基因学确诊NICCD患儿血清总胆红素、直接胆红素、谷草转氨酶、谷丙转氨酶及乳酸水平升高均占100%,血氨及甲胎蛋白水平升高均占95.2%,胆汁酸水平升高占90.0%,低蛋白水平占84.0%,血脂水平升高占50%,半乳糖增高占78.3%,4-羟基苯乳酸增高占52.2%,瓜氨酸增高占66.7%,C14增高占84.7%,C16增高占71.4%。结论：四个热点突变中851del4突变为此项研究病例中最常见的突变类型,其次为IVS6+5GA,然后是IVS16ins3kb和1638ins23。纯合突变、杂合突变之间患儿的瓜氨酸,酪氨酸,苯丙氨酸,C2,C14,C16浓度均无明显统计学差异。NICCD患儿平均3个月龄时体内存在明显的生化改变。NICCD患儿存在糖,氨基酸及脂肪酸代谢异常,并以半乳糖、瓜氨酸及长链酰基肉碱增高明显为主。
[Abstract]:Objective: to investigate the characteristics of common hot spot mutation gene and the relationship between homozygous mutation and heterozygous mutation in neonatal intrahepatic cholestasis (NICCD) caused by Citrin deficiency in Shenzhen area. To investigate the clinical manifestations and biochemical changes of children with NICCD and to provide theoretical basis for early diagnosis and treatment of children with NICCD. Methods: high resolution fusion curve (HRM) was used to screen four hot spot gene types (851del4p1638ins23, IVS6 5G > A and IVS16ins3kb) in children with clinically suspected NICCD. Gas chromatography-mass spectrometry (GC/MS) and tandem mass spectrometry (MS/MS) were used to detect the metabolites of carbohydrate, amino acid and fatty acid in urine and blood. Liver enzyme, blood ammonia, lactic acid, alpha-fetoprotein, lipid and other routine biochemical indicators and analysis. Results: a total of 27 cases were screened and diagnosed in 360 suspicious children, and the positive rate was 7.5%. Of the four hot spot mutations, 851del4 mutation accounted for 50.0%, 1638ins23% 13.0%, IVS16ins3kb 7.4kb, IVS6 5GA 7.4kb and IVS6 5GA 7.4kb. Citrulline, tyrosine, phenylalanine, acetyl carnitine (C2), myristic carnitine (C14) in children with homozygous mutation and heterozygous mutation. The serum total bilirubin, direct bilirubin, aspartate aminotransferase, alanine aminotransferase and lactic acid levels were 100% in 27 patients with NICCD. The levels of blood ammonia and alpha-fetoprotein were 95.2%, bile acid 90.0%, low protein 84.0%, lipids 50%, galactose 78.3%, serum ammonia 95.2%, bile acid 90.0%, low protein 84.0%, lipids 50%, galactose 78.3%. 4- hydroxyphenyllactic acid increased 52.2%, citrulline increased 66.7%, C14 increased 84.7% and C16 increased 71.4%. Conclusion: among the four hot spot mutations, 851del4 mutation is the most common mutation type, followed by IVS6 5GA, IVS16ins3kb and 1638ins2323. There was no significant difference in the concentrations of citrulline, tyrosine, phenylalanine, C2C14C16 between homozygous mutation and heterozygosity mutation. There were significant biochemical changes in the body of NICCD children at an average age of 3 months, and sugar in NICCD children. The metabolism of amino acids and fatty acids was abnormal, and the increase of galactose, citrulline and long chain acyl carnitine was significant.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R722.11

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