Child, Preschool ; Humans ; Male ; Mucopolysaccharidosis II ; diagnosis ; genetics ; therapy ; Siblings

OBJECTIVETo identify the mutations of iduronate-2-sulfatase (IDS) gene in mucopolysaccharidosis type II patients.

METHODSPCR-SSCP analysis was applied to detect the common mutations in the exons 2, 3, 5, 7, 8, 9 in IDS-gene of the patient. DNA sequencing and PCR-RFLP were applied to analyze the mutation detected by PCR-SSCP.

RESULTSA new mutation(1253G-->T) of exon 7 of the IDS gene was found by PCR-SSCP and DNA sequencing in the patient, The PCR-restriction enzyme digestion showed that enzyme digestion location appeared in the patient and his mother, which verified the results of sequencing analysis.

CONCLUSIONThe mutation of patient with MPSII could be detected effectively and quickly by the applications of PCR-SSCP, DNA sequencing and PCR-restriction enzyme digestion analysis, and the new mutation thus detected is necessary for the prenatal diagnosis of the pedigree.

Child ; Humans ; Iduronate Sulfatase ; genetics ; Male ; Mucopolysaccharidosis II ; genetics ; Mutation ; Polymorphism, Single-Stranded Conformational

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Male ; Mucopolysaccharidosis II ; genetics ; Pedigree

OBJECTIVE: To explore the genetic etiology of a patient with mucopolysaccharidosis type II (MPSII). METHODS: The IDS gene of the proband and his mother was detected by Sanger sequencing, agarose gel electrophoresis, real-time PCR and multiple ligation-dependent probe amplification (MLPA). Prenatal diagnosis was performed on amniotic fluid sample. RESULTS: Agarose gel electrophoresis, real-time PCR, and MLPA all showed that exon 2 of IDS gene of the proband was deleted, for which his mother was normal. Prenatal diagnosis showed that the fetus was a normal male. CONCLUSION The de novo deletion of exon 2 of the IDS gene probably underlay the MPSII in this patient. Above finding has broadened the mutation spectrum of the IDS gene. The combined methods for the detection of IDS gene mutations could make accurate prenatal diagnosis for MPSII.
China ; Exons ; Female ; Humans ; Male ; Mucopolysaccharidosis II/genetics* ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis/methods*

OBJECTIVETo identify the mutations of iduronate-2-sulfatase (IDS) gene, to reveal its mutation features, and to establish a basis for genetic counseling and prenatal gene diagnosis of Hunter syndrome.

METHODSUrine glycosaminoglycans (GAGs) assay, PCR and DNA sequencing were performed to detect mutation of IDS gene of the patient and his parents.

RESULTSThe result showed that the patient was: DS(++), HS(++), KS(-), CS(-), and that both of his parents were negative. A frame-shift deletion mutation (1062 del 16) was identified in exon 7 of the patient's IDS gene. His parents' genotypes were normal.

CONCLUSIONThe patient's mutation was not inherited by his parents but a novel one. The mutation probably altered the primary structure and tertiary structure of IDS enzyme protein remarkably and lowered the activity of IDS enzyme greatly. Therefore it is supposed to be the direct cause of the disorder.

Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child, Preschool ; Female ; Glycoproteins ; genetics ; urine ; Humans ; Male ; Mucopolysaccharidosis II ; enzymology ; genetics ; urine ; Mutation ; genetics

OBJECTIVETo identify the mutations of iduronate-2-sulfatase (IDS) gene, and to establish a basis of prenatal gene diagnosis of Hunter syndrome.

METHODSUrine glycosaminoglycan (GAG) assay was used to preliminary diagnosis of mucopolysaccharidosis. PCR-denaturing high-performance liquidchromatograptly (PCR-DHPLC) analysis was performed to detect the mutation in exons 9, 3, 8 of the IDS gene. DNA sequencing was applied to analyze the mutation detected by PCR-DHPLC.

RESULTSAbnormal peaks were found by PCR-DHPLC. A new frame-mutation (1569+TT) in exon 9 of IDS gene was identified by DNA sequencing. Two "T"q inserted in position 1569 base pair (1569+TT) caused a substitution of codon 482 (TTA, leucine) to 482 (TTT, phenylalanine). The "TT" insertion results in the decrease of amino acids from 550 to 482. The patient is a hemizygote and his mother is a heterozygote.

CONCLUSIONA new frame-shift mutation of IDS gene is found to report. The mutation (1569+TT) results in 68 amino acids lost. Probably it causes the enzyme activity seriously dropped down and being pathologically the basis of disease.

Child, Preschool ; Chromatography, High Pressure Liquid ; DNA Mutational Analysis ; Humans ; Iduronate Sulfatase ; genetics ; Male ; Molecular Sequence Data ; Mucopolysaccharidoses ; genetics ; Mucopolysaccharidosis II ; enzymology ; genetics ; Mutation

OBJECTIVETo establish a method of iduronate-2-sulfatase (IDS) activity assay and mutation analysis of IDS gene for the prenatal diagnosis of mucopolysaccharidosis type II (MPSII).

METHODSPrenatal diagnosis of two cases was performed using cultured fetal amniotic fluid cells. Enzyme activity of IDS in cultured fetal amniotic fluid cells extracted from the two pregnant women at high risk of MPS II was measured. Meanwhile, genomic DNA was extracted for fetal gender testing and mutation analysis of the IDS gene.

RESULTSEnzyme activity assay showed that IDS activity in amniotic fluid cells was significantly decreased. IDS gene sequencing showed that the male fetus was hemizygous mutant, and the female fetus was carrier of heterozygous mutation. Therefore the male fetus was an MPS II patient and the female fetus was a mutation carrier.

CONCLUSIONDetermination of IDS activity in fetal amniotic fluid cells together with IDS gene mutation analysis is a rapid, sensitive and accurate method of prenatal diagnosis of MPS II. Using this method, prenatal diagnosis for pregnant women at high risk of MPSII can be achieved.

Base Sequence ; Child ; Child, Preschool ; Exons ; Female ; Glycoproteins ; genetics ; metabolism ; Humans ; Male ; Mucopolysaccharidosis II ; diagnosis ; genetics ; metabolism ; Mutation ; genetics ; Pregnancy ; Prenatal Diagnosis

OBJECTIVETo detect potential mutation of iduronate-2-sulfatase (IDS) gene in a family affected with mucopolysaccharidosis type Ⅱ (MPS Ⅱ).

METHODSFor the proband and his unaffected mother, the whole coding sequence of the IDS gene was analyzed with PCR and bidirectional Sanger sequencing.

RESULTSA novel splicing mutation, c.709-1G>A, was detected in the proband, for which his mother was heterozygous.

CONCLUSIONThe c.709-1G>A splicing mutation of the IDS gene is probably causative for the MSP Ⅱ in the proband. Prenatal diagnosis for the mutation may avoid birth of further child affected with this disease.

Base Sequence ; Child ; DNA Mutational Analysis ; methods ; Family Health ; Female ; Genetic Predisposition to Disease ; genetics ; Glycoproteins ; genetics ; metabolism ; Heterozygote ; Humans ; Iduronate Sulfatase ; genetics ; metabolism ; Male ; Mothers ; Mucopolysaccharidosis II ; diagnosis ; enzymology ; genetics ; Mutation

OBJECTIVEMucopolysaccharidosis type II (MPSII) is a lethal, X-linked recessive disorder caused by mutation of iduronate-2-sulfatase (IDS) gene. Up to now there is no really effective treatment for this disorder, therefore it is important to provide an accurate genetic diagnosis and prenatal diagnosis for the MPSII families. In this study, we identify the pathogenic mutation in a Chinese family with MPSII.

METHODThe 8 years old male proband from a Chinese family was clinically diagnosed with MPSII. There are other 4 patients with similar phenotypes in the family who died at 9, 11, 7 and 10 years of age, respectively. Mutation analysis was carried out by polymerase chain reaction and direct sequencing of all exons and exon/intron boundaries of IDS gene. Denaturing high performance liquid chromatography (DHPLC) analysis was performed to screen the unknown variations of IDS gene in 100 unrelated control males.

RESULTTwo allelic variants of exon 5 (c.684A > G) and exon 6 (c.851C > T) and a nonsense mutation of exon 7 (c.892C > T) were detected in IDS gene of the proband. Heterozygous mutations c.684A > G, c.851C > T and c.892C > T were detected in both proband's mother and maternal grandmother. The unknown variations of c.684A > G and c.851C > T were not found in the 100 unrelated control males. The male fetus (IV11) inherited the same mutation of IDS gene as the proband.

CONCLUSIONMutation c.892C > T of IDS gene causes MPSII in this family and prenatal diagnosis in one affected fetus was achieved.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Child ; DNA Mutational Analysis ; Family ; Female ; Humans ; Iduronate Sulfatase ; genetics ; Male ; Middle Aged ; Mucopolysaccharidosis II ; diagnosis ; genetics ; Mutation ; Phenotype ; Pregnancy ; Prenatal Diagnosis

OBJECTIVETo investigate and establish the gene diagnosis methods for the frequent mutations of iduronate-2-sulphatase(IDS) gene in mucopolysaccharidosis type II patients.

METHODSpolymerase chain- reaction-single strand conformation polymorphism PCR-SSCP) analysis was applied to detect the mutations of exons 3, 8 and 9 which were hot spots in the iduronate-2-sulfatase gene; DNA sequencing was applied to analyze the mutations which had been detected by PCR-SSCP; PCR-restriction fragment length polymorphism (PCR-RFLP) was applied to detect the results of DNA sequencing.

RESULTSObvious and abnormal bands in exon 9 of the IDS gene were found by applying PCR-SSCP; the mutation(C1672T) of exon 9 was found in the patient through DNA sequencing, which led to amino acid replacement(R468W); the PCR-restriction enzyme digestion showed that only one band(554 bp) appeared in the patient, but there were two bands (257 bp and 297 bp) in his parents, and it verified the results of sequencing analysis.

CONCLUSIONPCR-SSCP analysis, DNA sequencing analysis and PCR-restriction enzyme digestion are effective methods for MPS II diagnosis. Combined applications of these methods can verify and complement each other and improve the accuracy of diagnosis.

Amino Acid Substitution ; Base Sequence ; Child ; China ; Codon, Nonsense ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Humans ; Iduronate Sulfatase ; genetics ; Male ; Mucopolysaccharidosis II ; enzymology ; genetics ; Mutation ; Point Mutation ; Polymorphism, Single-Stranded Conformational