21-羟化酶缺乏症基因型和临床表型分析
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摘要
目的了解21-羟化酶缺乏症(21-OHD)患儿CYP21A2基因突变热点,并分析其基因型和表型的关系。方法对在新生儿17α羟孕酮(17-αOHP)筛查中发现并诊断的18例21-OHD患儿及其父母,采用等位基因特异性PCR-直接测序联合多重连接依赖探针扩增技术测定其基因型。结果共检出6种CYP21A2基因突变类型,最常见的为I2G(44.4%)和de(l33.3%),这两种也是失盐型常见的突变;单纯男性化型患儿I172N突变检出率达75%。根据对21-羟化酶活性影响程度将患儿分为重度、中度和轻度组,三组间反映病情严重性的指标17-αOHP、促肾上腺皮质激素、睾酮的差异均有统计学意义(P<0.05)。结论 21-OHD基因诊断技术的建立揭示了基因型和临床表型的一致性。
Objectives To study the mutation spectrum in CYP21A2 gene in patients with 21-hydroxylase deficiency(21-OHD),and to analyze the relationship between genotype and phenotype.Methods Eighteen patients with 21-OHD were identified by neonatal screening of 17α-OH progesterone(17α-OHP).The allele specific PCR-DNA sequencing combining with multiplex ligation-dependent probe amplification was applied to determine the genotype in the patients and their parents.Results Six mutations of CYP21A2 gene were identified.I2G(44.4%) and del(33.3%) were the most frequent mutations and also were the most common mutations in salt-wasting form.The detection rate of I172N mutation in simple virilizing form was 75%.Patients were classified into three groups according to the degree of 21-hydroxylase enzymatic compromise caused by the mutation.The serum 17α-OHP,ACTH and T levels which reflected the severity of disease were significantly different among three groups(P<0.05).Conclusions The genetic diagnosis of 21-OHD reveals the consistency between genotype and phenotype.
Objectives To study the mutation spectrum in CYP21A2 gene in patients with 21-hydroxylase deficiency(21-OHD),and to analyze the relationship between genotype and phenotype.Methods Eighteen patients with 21-OHD were identified by neonatal screening of 17α-OH progesterone(17α-OHP).The allele specific PCR-DNA sequencing combining with multiplex ligation-dependent probe amplification was applied to determine the genotype in the patients and their parents.Results Six mutations of CYP21A2 gene were identified.I2G(44.4%) and del(33.3%) were the most frequent mutations and also were the most common mutations in salt-wasting form.The detection rate of I172N mutation in simple virilizing form was 75%.Patients were classified into three groups according to the degree of 21-hydroxylase enzymatic compromise caused by the mutation.The serum 17α-OHP,ACTH and T levels which reflected the severity of disease were significantly different among three groups(P<0.05).Conclusions The genetic diagnosis of 21-OHD reveals the consistency between genotype and phenotype.
引文
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[12]Bas F,Kayserili H,Darendeliler F,et al.CYP21A2 gene mu-tations in congenital adrenal hyperplasia:genotype-phenotypecorrelation in Turkish children[J].J Clin Res Pediatr Endocri-nol,2009,1(3):116-128.
[13]Shinagawa T,Hofikawa R,Isojima T,et a1.Nonclassicsteroid 21-hydroxylase deficiency due to a homozygous V281L.mutation in CYP21A2 detected by the neonatal mass-screeningprogram in Japan[J].Endocr J,2007,54(6):1021-1025.
[14]Concolino P,Mello E,Patrosso MC.p.H282N and p.Y191H:2 novel CYP21A2 mutations in Italian congenital adrenal hy-perplasia patients[J].Metabolism,2012,61(4):519-524.
[15]Baumgartner-Parzer S M,Fischer G,et al.Predisposition forde novo gene aberrations in the offspring of mothers with aduplicated CYP21A2 gene[J].J Clin Endocrinol Metab,2007,92(3):1164-1167.
[16]Marumudi E,Sharma A,Kulshreshtha B,et al.Moleculargenetic analysis of CYP21A2 gene in patients with congenitaladrenal hyperplasia[J].Indian J Endocrinol Metab,2012,16(3):384-388.
[17]Oriola J,Bosch MZ,Valls C,et al.Association ofp.His38Leu,a rare CYP21A2 mutation,with the classicalsimple virilizing phenotype of 21-hydroxylase deficiency in a6-year-old boy[J].Horm Res Paediatr,2011,76(3):214-217.
[2]Balsamo A,Cacciari E,Baldazzi L,et al.CYP21 analysisand phenotype/genotype relationship in the screened popula-tion of the Italian Emilia-Romagna region[J].Clin Endocrinol,2000,53(1):117-125.
[3]张波,陆召麟,王玥.中国人21羟化酶缺乏症基因型和临床表型特点研究[J].遗传学报,2004,31(9):950-959.
[4]Tsai LP,Lee HH.Analysis of CYP21A1P and the duplicatedCYP21A2 genes[J].Gene,2012,506(1):261-262.
[5]Riepe FG,Sippell WG.Recent advance in diagnosis treatmentand outcome of congenital adrenal hyperplasia due to 21-hy-droxylase deficiency[J].Rev Endocr Metab Disord,2007,8(4):349-363.
[6]Keen-Kim D,Redman JB,Alanes RU,et al.Validation andclinical application of a locus-specific polymerase chain reac-tion and minisequencing-based assay for congenital adrenalhyperplasia(21-hydroxylase deficiency)[J].J Mol Diagn,2005,7(2):236-246.
[7]Concolino P,Mello E,Toscano V,et al.Multiplex ligation-dependent probe amplification(MLPA)assay for the detectionof CYP21A2 gene deletions/duplications in congenital adrenalhyperplasia:first technical report[J].Clin Chim Acta,2009,402(1-2):164-170.
[8]Schouten JP,McElgunn CJ,Waaijer R,et al.Relative quan-tification of 40 nucleic acid equences by multiplex ligation-dependent probe amplication[J].Nucleic Acids Res,2002,30(12):e57.
[9]Witchel SF,Smith R,Crivellaro CE,et al.CYP21 muta-tions in Brazilian patients with 21-hydroxylase deficiency[J].Hum Genet,2000,106(4):414-419.
[10]郁婷婷,王剑,余永国,等.先天性肾上腺皮质增生症基因突变位点诊断方法的建立[J].中华检验医学杂志,2010,33(7):606-610.
[11]Chen W,Xu Z,Nishitani M,et al.Complement component4 copy number variation and CYP21A2 genotype associationsin patients with congenital adrenal hyperplasia due to 21-hy-droxylase deficiency[J].Hum Genet,2012,131(12):1889-1894.
[12]Bas F,Kayserili H,Darendeliler F,et al.CYP21A2 gene mu-tations in congenital adrenal hyperplasia:genotype-phenotypecorrelation in Turkish children[J].J Clin Res Pediatr Endocri-nol,2009,1(3):116-128.
[13]Shinagawa T,Hofikawa R,Isojima T,et a1.Nonclassicsteroid 21-hydroxylase deficiency due to a homozygous V281L.mutation in CYP21A2 detected by the neonatal mass-screeningprogram in Japan[J].Endocr J,2007,54(6):1021-1025.
[14]Concolino P,Mello E,Patrosso MC.p.H282N and p.Y191H:2 novel CYP21A2 mutations in Italian congenital adrenal hy-perplasia patients[J].Metabolism,2012,61(4):519-524.
[15]Baumgartner-Parzer S M,Fischer G,et al.Predisposition forde novo gene aberrations in the offspring of mothers with aduplicated CYP21A2 gene[J].J Clin Endocrinol Metab,2007,92(3):1164-1167.
[16]Marumudi E,Sharma A,Kulshreshtha B,et al.Moleculargenetic analysis of CYP21A2 gene in patients with congenitaladrenal hyperplasia[J].Indian J Endocrinol Metab,2012,16(3):384-388.
[17]Oriola J,Bosch MZ,Valls C,et al.Association ofp.His38Leu,a rare CYP21A2 mutation,with the classicalsimple virilizing phenotype of 21-hydroxylase deficiency in a6-year-old boy[J].Horm Res Paediatr,2011,76(3):214-217.