Předmětem projektu je zdokonalení současného stavu DNA diagnostiky kongenitální adrenální hyperplasie (CAH) v České republice. Studiem genů CYP21 a CYP11B1 s využitím technik jako MLPA, LR PCR, gene walking, DHPLC a sekvenování chceme zvýšit efektivitu mutační analýzy CAH v rodinách s výskytem tohoto život ohrožujícího metabolického onemocnění.; The aim of the project is to improve present DNA diagnostics of congenital adrenal hyperplasia (CAH) in the Czech Republic. By study of CYP21 and CYP21P genes with using of techniques such as MLPA, LR PCR, gene walking, DHPLC and sequencing we want to increase the effectiveness of mutation analysis of CAH in affected families.

• MeSH
• Molecular Diagnostic Techniques MeSH
• Adrenal Hyperplasia, Congenital MeSH
• DNA Mutational Analysis MeSH
• Steroid 21-Hydroxylase MeSH
• Steroid Hydroxylases MeSH
• Metabolism, Inborn Errors MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• endokrinologie
• genetika, lékařská genetika
• biologie
• NML Publication type
• závěrečné zprávy o řešení grantu IGA MZ ČR

Předmětem projektu je zdokonalení současného stavu DNA diagnostiky v České republice. Metodou APEX s využitím DNA čipů (microarrays) chceme zvýšit efektivitu mutační analýzy u dědičných onemocnění.; The aim of the project is to improve present DNA diagnosis in the Czech Republic. We want to increase the effectiveness of mutation analysis of inherited disorders by the introduction of the APEX microarray method.

• MeSH
• Molecular Diagnostic Techniques MeSH
• Genetic Diseases, Inborn diagnosis   MeSH
• Microarray Analysis MeSH
• Mutation MeSH
• Oligonucleotide Array Sequence Analysis MeSH
• Metabolism, Inborn Errors diagnosis   MeSH

• Conspectus
• Biochemie. Molekulární biologie. Biofyzika
• NML Fields
• biologie
• genetika, lékařská genetika
• NML Publication type
• závěrečné zprávy o řešení grantu IGA MZ ČR

• Publication type
• Abstracts MeSH

Wilson disease (WD) is an autosomal recessive disorder of copper transport. WD patients are presenting with a wide range of heterogeneous clinical syndromes including hepatic, neurological, or psychiatric presentations. The disease is caused by mutations in the ATP7B gene. This study presents the results of comprehensive mutation analysis in 227 WD patients from 200 unrelated families (173 from Czech Republic and 27 from Slovakia). More than 80% of all mutant alleles were identified, using a combination of PCR/RFLP, DGGE, TTGE, DHPLC, and sequencing. A total of 40 different mutations and 18 polymorphisms were detected on 400 independent mutant chromosomes. The most common molecular defect was H1069Q (57% of all 400 studied alleles). Each of the other 39 mutations was present in no more than 4% of WD alleles and 23 mutations were found in only one WD allele each (0.25%). Thirteen novel mutations were identified, including seven missense mutations (L641S, T737R, D918E, T1033S, G1111D, D1271N, and G1355C), four small deletions (19_20delCA, 1518_1522del5, 3140delA, and 3794_3803del10), and two splice-site mutations (2446-2A>G, 2865+1G>A). We did not find a significant correlation between H1069Q homozygosity and age of onset, and clinical and biochemical manifestation. Our data provide evidence that the H1069Q mutation-the most common molecular defect of the ATP7B gene in the Caucasian population-originates from Central/Eastern Europe. Screening of five prevalent mutations is predicted to reveal 70% of all mutant alleles presented in WD patients. This will provide a good starting point for early clinical classification of WD in our population.

• MeSH
• Adenosine Triphosphatases genetics   MeSH
• DNA Primers MeSH
• Electrophoresis, Polyacrylamide Gel MeSH
• Phenotype MeSH
• Financing, Organized MeSH
• Genotype MeSH
• Hepatolenticular Degeneration genetics   MeSH
• Humans MeSH
• Mutation MeSH
• Polymerase Chain Reaction MeSH
• Polymorphism, Restriction Fragment Length MeSH
• Cation Transport Proteins genetics   MeSH
• Base Sequence MeSH
• Check Tag
• Humans MeSH

Východisko. Wilsonova choroba je autozomálně recesivní onemocnění charakteristické akumulací a intoxikací mědi v organizmu. Molekulární podstatou onemocnění jsou mutace v genu pro měď transportující ATPázu (ATP7B). Metody a výsledky. Předmětem sdělení jsou výsledky molekulárně-genetických vyšetření ve 130 nepříbuzných rodinách s výskytem Wilsonovy choroby. Pomocí denaturační gradientové gelové elektroforézy (DGGE) byly detekovány exony s abnormální sekvencí. Následným sekvenováním bylo nalezeno celkem 17 kauzálních mutací a 9 polymorfizmů. Pět mutací bylo dosud nepopsaných. Z celkového počtu 260 mutantních alel bylo 214 (82,3 %) identifikováno. Nejčastější mutace, H1069Q, se v naší populaci vyskytuje s frekvencí 65,8 %. Frekvence ostatních mutací nepřevyšuje 5 %. Závěry. DNA diagnostika Wilsonovy choroby v postižených rodinách nabízí pohotové a spolehlivé výsledky. V rámci genetického poradenství umožňuje odhalit asymptomatické a heterozygotní sourozence.

Background. Wilson disease is an autosomal recessive disorder, characterized by cooper accumulation and intoxication of the organism. Molecular basis of the disease represent mutations in the gene for the copper-transporting ATPase (ATP7B). Methods and Results. The submitted paper deals with results of molecular-genetic examination in 130 unrelated families in whichWilson disease was diagnosed. By denaturing gradient gel electrophoresis (DGGE), the exons with abnormal sequences were detected. Followed by sequencing, 17 causal mutations and 9 silent polymorphism were found. Five novel mutations were detected. After analysis of 260 mutant alleles, 214 (82.3 %) were identified. The most frequent mutation, H1069Q, occurred in our population with the frequency of 65.8 %. Incidence of other mutations, however, did not exceed 5 %. Conclusions. DNA analysis of the Wilson disease offers prompt and reliable results in affected families. It can help to identify asymptomatic and heterozygote siblings at genetic counselling.

• MeSH
• Adenosine Triphosphatases MeSH
• Child MeSH
• Adult MeSH
• Research Support as Topic MeSH
• Genes MeSH
• Hepatolenticular Degeneration diagnosis   etiology   MeSH
• Humans MeSH
• Copper metabolism   urine   MeSH
• Molecular Biology methods   MeSH
• Mutation MeSH
• DNA Mutational Analysis MeSH
• Check Tag
• Child MeSH
• Adult MeSH
• Humans MeSH
• Publication type
• Review MeSH
• Comparative Study MeSH