[A novel mutation causes congenital factor V deficiency].
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To investigate the gene defect in a hereditary coagulation factor V (FV) deficiency family.The plasma FV actigen was measured by one-stage clotting assay. The FV antigen was assayed by Biotin-Avidin enzyme linked immunosorbent assay (BA-ELISA). The full length of exon 1 to exon 25 and the 5' untranslated sequence of FV genomic DNA were analyzed by polymerase chain reaction (PCR) and direct sequencing of the amplified fragments, meanwhile the defect was identified by T/A cloning sequencing.The plasma coagulant activity and amount of FV of the proband were marked deficient (1% and 1.54%, respectively). DNA sequence analysis for the proband revealed a causative mutation in a heterozygous status. It was one base pair deletion in exon 4 at nucleotide 675 inherited from her mother.A novel mutation in the FV gene was identified in the proband with congenital FV deficiency. The mutation was 675delA in exon 4 resulting in a frameshift and a premature termination codon.Keywords:
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[Identification of a novel EXT1 mutation in a pedigree affected with hereditary multiple exostosis].
OBJECTIVE To detect potential mutations of the EXT1 and EXT2 genes in a pedigree affected with hereditary multiple exostosis (HME). METHODS For a four-generation family with 7 affected individuals from 17 family members,genomic DNA was extracted from peripheral venous blood samples. All exons of the EXT1 and EXT2 genes were screened for potential mutation by PCR and Sanger sequencing. RESULTS A novel heterozygous frameshift mutation c.1202delT (p.I401Tfs*2)was found in exon 4 of the EXT1 gene in the proband and the other 6 affected individuals. The same mutation was not detected among the healthy members from the family. The mutation has given rise a truncated EXT1 protein with loss of 345 amino acids. CONCLUSION A novel frameshift mutation of the EXT1 gene has been identified in a pedigree affected with HME, which has enriched the mutational spectrum of the EXT1 gene and may facilitate genetic counseling and prenatal diagnosis for the family.
Hereditary multiple exostoses
Sanger sequencing
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genomic DNA
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To detect the underlying genetic defect in two Chinese families with hereditary multiple exostoses and provide genetic counseling.Potential mutations in EXT1 and EXT2 genes in the probands were detected by direct sequencing of PCR-amplified exons. Suspected mutations were verified in all available family members and 200 unrelated healthy controls.A heterozygous frameshift mutation c.346_356delinsTAT in exon 1 of EXT1 and a heterozygous deletion mutation c.2009-2012del(TCAA) in exon 10 of EXT1 were respectively detected in affected members from the two families. The same mutations were not detected in unaffected members and 200 unrelated healthy controls. No mutations in EXT2 were detected in the two families.Two novel mutations of EXT1 have been detected in association with hereditary multiple exostoses in two Chinese families. Above results have provided a basis for genetic counseling for the two families and expanded the spectrum of EXT1 mutations.
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Abstract Background: Holocarboxylase synthetase (HLCS) deficiency is a rare inborn disorder of biotin metabolism, which results in the defect of several biotin-dependent carboxylases and presents with metabolic ketoacidosis and skin lesions. Case presentation : In this study, we have reported a Chinese Han pedigree with HLCS deficiency diagnosed using next-generation sequencing and validated with Sanger sequencing of HLCS and BTD gene. The Chinese proband carries a common missense mutation c.1522C>T (p.Arg508Trp) in exon 9 of HLCS gene, which generates an increased K m for biotin. A novel frameshift mutation c.1006_1007delGA (p.Glu336Thrfs*15) in exon 6 of H L CS gene produces a low Vmax and is predicted to be deleterious through PROVEAN and MutationTaster. A novel heterozygous mutation c.638_642delAACAC (p.His213Profs*4) in BTD gene is also identified. Otherwise, the proband presents abnormal BAEP suggesting hearing damage in the acute episode. Conclusions: A Chinese proband carries a reported Arg508Trp variant, a novel 2-bp frameshift mutation c.1006_1007delGA (p.Glu336Thr) expanding mutational spectrum of HLCS gene, and a novel heterozygous mutation c.638_642delAACAC (p.His213Profs*4) expanding mutational spectrum of BTD gene. Furthermore, the reversible hearing damage is rarely reported in the patients with HLCS deficiency, which deserves for further discussion.
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Compound heterozygosity
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To investigate the gene defect in a hereditary coagulation factor V (FV) deficiency family.The plasma FV actigen was measured by one-stage clotting assay. The FV antigen was assayed by Biotin-Avidin enzyme linked immunosorbent assay (BA-ELISA). The full length of exon 1 to exon 25 and the 5' untranslated sequence of FV genomic DNA were analyzed by polymerase chain reaction (PCR) and direct sequencing of the amplified fragments, meanwhile the defect was identified by T/A cloning sequencing.The plasma coagulant activity and amount of FV of the proband were marked deficient (1% and 1.54%, respectively). DNA sequence analysis for the proband revealed a causative mutation in a heterozygous status. It was one base pair deletion in exon 4 at nucleotide 675 inherited from her mother.A novel mutation in the FV gene was identified in the proband with congenital FV deficiency. The mutation was 675delA in exon 4 resulting in a frameshift and a premature termination codon.
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genomic DNA
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Multiple osteochondromas (MO), an inherited autosomal dominant disorder, is characterized by the presence of multiple exostoses on the long bones. MO is caused by mutations in the EXT1 or EXT2 genes which encode glycosyltransferases implicated in heparin sulfate biosynthesis.In this study, efforts were made to identify the underlying disease-causing mutations in patients from two MO families in China.Two novel EXT1 gene mutations were identified and no mutation was found in EXT2 gene. The mutation c.497T > A in exon 1 of the EXT1 gene was cosegregated with the disease phenotype in family 1 and formed a stop codon at amino acid site 166. The fetus of the proband was diagnosed negative. In family 2, the mutation c.1430-1431delCC in exon 6 of the EXT1 gene would cause frameshift and introduce a premature stop codon after the reading frame being open for 42 amino acids. The fetus of this family inherited this mutation from the father.Mutation analysis of two MO families in this study demonstrates its further application in MO genetic counseling and prenatal diagnosis.
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Hereditary multiple exostoses
Stop codon
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AIM:To find potential mutable sites by detecting mutations of the candidate gene in a kindred with polycystic liver disease (PCLD). METHODS:First, we chose a kindred with PCLD and obtained five venous blood samples of this kindred after the family members signed the informed consent form.In the kindred two cases were diagnosed with PCLD, and the left three cases were normal individuals.All the blood samples were preserved at -85 ℃.Second, we extracted the genomic DNA from the venous blood samples of the kindred using a QIAamp DNA Mini Kit and then performed long-range polymerase chain reaction (PCR) with different primers.The exons of sequence.This mutation was located in the first codon and resulted in a termination codon.This mutation had an obvious influence on the encoded protein and changed the length of the protein from 4303 to 2246 amino acids.This was a new mutation that was not present in the dbSNP library. CONCLUSION:The nonsense mutation of exon 15 existed in the proband and in the third individual.Additionally, the proband was heterozygous for this mutation, so the mutable site was a pathogenic mutation.
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Nonsense mutation
genomic DNA
Stop codon
Silent mutation
Sanger sequencing
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Objective To investigate the EXT gene mutation in a pedigree with hereditary multiple exostoses(HME).Methods All exons and their flank sequences in EXT1 and EXT2 genes obtained from proband’s genomic DNA were amplified by PCR,the PCR products were then purified and directly sequenced.Results A heterozygous mutation,1564-7delC was found in proband’s EXT1 gene.The mutation caused a frameshift from 522-amino acid site and a premature terminating code was introduced at 546-amino acid site,producing a truncated protein.Family pedigree study showed that this mutation was come from proband’s mother.No variant was found in EXT2 gene.Conclusions 1564-7delC heterozygous mutation in EXT1 gene is the molecular mechanism of developing HME in this pedigree.
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Hereditary multiple exostoses
genomic DNA
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Objective To identify the mutation of DSRAD gene in patients with DSH. Methods Blood samples were collected from the patients and healthy members of a DSH family and 100 unrelated controls. All 15 exons of the DSRAD gene were analyzed by PCR-DNA sequencing. Results A frameshift mutation,c.1615delG in the exon 3 of the DSRAD gene,was found in the affected members but not in the healthy individuals of this family and 100 unrelated controls. Conclusion There is a novel frameshift mutation in DSRAD gene in this DSH family.
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OBJECTIVE To detect potential mutation in a Chinese family affected with beta-ureidopropinoase deficiency. METHODS Genomic DNA was extracted from peripheral blood samples. All exons and flanking intron regions of the UPB1 gene were amplified by PCR and detected by direct sequencing. RESULTS A homozygous mutation c.977G>A was identified in exon 9 of the UPB1 gene in the proband. Both parents of the proband had heterozygous change of the same site. CONCLUSION The c.977G>A mutation of the UPB1 gene is responsible for the pathogenesis of the disease in the infant.
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genomic DNA
Pathogenesis
Mutation Testing
BETA (programming language)
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Objective To investigate EXT gene sequence in a pedigree with hereditary multiple exostoses (HME) and identify the causative mutation in the pedigree. Methods All the exons and their flanking sequences of EXT1 and EXT2 genes were amplified by PCR from proband′ s genomic DNA, PCR products were purified and sent for direct sequencing. Results A novel insertion-deletion (651 - 664 delins TTT) mutation in exon 1 of EXT1 gene was found, the mutation resulted in a frameshift at amino acid position 218 and a premature stop codon at position 220 (K218 fs X220) in the EXT1 protein. The mutation also lead to a truncated protein. Family study results showed that this mutation came from the proband′s mother. No variant was found in EXT2 gene of this proband. Conclusions 651 - 664 delins TTT heterozygous mutation in EXT1 gene was the molecular mechanism of HME for this pedigree.
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Hereditary multiple exostoses
genomic DNA
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