黏多糖贮积症Ⅲ型的鉴别诊断与产前诊断
张为民;施惠平;孟岩;李贝特;邱正庆;刘俊涛
【期刊名称】《中华儿科杂志》 【年(卷),期】2008(046)006
【摘要】Objective Mucopolysaceharidosis(MPs)types ⅢA,B,C,D are a group of autosomal recessive lysosomal storage disorders caused by mutations in one of four genes which encode enzyme activities reqtIired for the lysosomal degradation of heparan sulfate.MPSⅢA and MPSⅢB involve
deficiencies
of
heparan
N-sulfatase(SGSH)and
α-N-
acetylglucosaminidase(NAGLU).MPSⅢA and MPSⅢB are more common than MPSⅢC and Ⅲ D.The present study aimed to establish two enzyme assay methods for SGSHand NAGLU activities for carrying out postnatal and prenatal diagnosis of MPSⅢA and ⅢB by means of SGSH and NAGLU activity assay on plasma,leukocyte,uncultured chorionic villi(CV) and cultured amniotic fluid cells(AF eell)using two newly synthesized substrates.Mutation analysis of SGSH gene was also performed.Methods Two fluorigenic substrate(4-methylumbelliferyl-α-D-N-sulphoglucosaminide.Na
and
4-methylumbelliferyl-α-N-
acetylglueosaminide)were used for the assay of SGSH and NAGLU activity.SGSH activitv in leukocyte was determined for diagnosis MPSⅢA proband.NAGLU activity was determinedin plasma for diagnosis of MPSⅢB proband.Twelve cases with MPS Ⅲ were enrolled
in this study,4 were female and 8 were male.age 3-10 years and were from 10 unrelated families.Eight exons of SGSH gene were amolified by PCR The mutations of the patients were characterized by direct sequencing of theamplified DNA fragments.Prenatal diagnosis in 3 pregnancies at risk was carried out according to NAGLU activitv on uncultured CV at 11th week or on cultured AF cell at 18th week of gestation.Results The respectively.Five cases of MPSⅢB and 7 cases of MPSⅢA were diagnosed.The mutation analysis of SGSH gene showed 6 mutations(G191R,D235N,R377C,E447K,R233X and D219Wfs264X),only oneof which (D219Wlfs264X) has not been previously reported.Prenatal diagnosis Was performed on 3Conclusions The method using synthesized
fluorigenic
4-methylumbelliferyl-substrates
were
sensitive,rapid and convenient assay of SGSH and NAGLU activity and were reliable for early prenatal diagnosis.Mutation analysis on MPSⅢA patients suggests new possibilities for molecular diagnosis of the disease.%目的 建立以人工合成荧光底物测定乙酰肝素N-硫酸酯酶(SGSH),α-N-乙酰葡萄糖苷酶(NAGLU)活性的方法,应用这两种方法对黏多糖贮积症Ⅲ型中的A、B亚型(MPSⅢA和ⅢB)病例进行鉴别诊断,并对MPSⅢB高危妊娠的孕妇进行了产前诊断,同时从分子遗传学方面对MPSⅢA患儿进行了SGSH基因分析.方法 采用人工合成的荧光底物(4-甲基伞形酮-α-D-N-硫酸葡萄糖苷4-methylumbelliferyl-α-D-N-sulphoglucosaminide.Na,4-甲基伞形酮-N-乙酰-α-葡萄糖苷4-methylumbelliferyl-α-N-acetylglucosaminide)测定疑似
患儿白细胞和血浆中SGSH及NAGLU活性,在收集的12例患儿中,女4例,男8例,年龄3~10岁.来自10个无相关的家庭.用MPSⅢA的患儿外周血白细胞DNA进行SGSH基因的外显子PCR扩增,并通过的序列测定确定突变位置.用绒毛或经培养的羊水细胞为检材,对MPSⅢB高危妊娠的孕妇进行产前诊断.结果 获得中国正常人白细胞中SGSH活性正常值4.4～8.1 nmot/(17 h·mg蛋白),各种组织中NAGLU活性正常值:血浆中为33.3～62.4 nmoL/(4 h·ml),绒毛组织为44.9~91.7 nmol/(17 h ·mg蛋白),经培养的羊水细胞为53.2-82.2 nmol/(17 h·mg蛋白);在12例疑似患儿中确诊了7例MPSⅢA(其中两例为同胞),5例MPS ⅢB患者(其中2例为同胞),SGSH基因分析发现6种突变类型,已知突变5种(G191R,D235N,R377C,E447K和R233X),插入突变1种(D219Wfs264X),此插入突变为新生突变.3例MPSⅢB产前诊断中,2例胎儿正常,1例胎儿受累.结论 应用荧光法测定SGSH和NAGLU酶活性是敏感、可靠、快速的方法,可用于MPSⅢA和ⅢB病例的鉴别诊断及产前诊断;SGSH基因分析方法成本低,突变检出率高,可用于MPSⅢA的诊断和产前诊断. 【总页数】4页(407-410)
【关键词】黏多糖累积病Ⅲ型;诊断,鉴别;产前诊断;突变 【作者】张为民;施惠平;孟岩;李贝特;邱正庆;刘俊涛
【作者单位】100730,中国医学科学院,中国协和医科大学,北京协和医院中心实验室;中国医学科学院,中国协和医科大学,基础医学研究所医学遗传学系;中国医学科学院,中国协和医科大学,基础医学研究所医学遗传学系;100730,中国医学科学院,中国协和医科大学,北京协和医院中心实验室;100730,中国医学科学院,中国协和医科大学,北京协和医院儿科;100730,中国医学科学院,中国协和医科大学,
北京协和医院妇产科 【正文语种】中文 【中图分类】R71 【文献来源】
https://www.zhangqiaokeyan.com/academic-journal-cn_chinese-journal-pediatrics_thesis/0201236150327.html 【相关文献】
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黏多糖贮积症Ⅲ型的鉴别诊断与产前诊断
