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Utilization of new generation sequencing methods to elucidate cystic fibrosis-like phenotype at patients with unclear illness of molecular type.
Matějčková, Iva ; Macek, Milan (advisor) ; Holá, Dana (referee)
Cystic fibrosis (CF) is genetically conditioned, autosomal recessive disease that occurs in the European population with a prevalence of about 1:2500 - 1:1800. In this disease we observe a mutation of the CTFR gene with subsequent fault in chloride channels. Such afflicted individuals usually suffer from chronic respiratory problems, pancreatic insufficiency, high concentration of chloride ions in sweat and obstructive azoospermia. Genetic testing of CFTR gene is indicated in individuals who meet the CF clinical picture and a positive sweat test (increased concentration of chlorides in the sweat). Genetic testing of the CFTR gene is usually done by using commercial kits detecting the most common mutations of the CFTR gene in the Czech Republic. If the testing results are negative, it is further performed an MLPA method that captures the larger deletions and duplications of gene, eventually a sequencing of all exons is. Despite the well-established algorithm of the testing, some patients suffering from symptoms of CF are left without genetic findings. Thanks to development of next generation sequencing, it is possible to make the diagnosis of CF more effective and uncover the variants that were not captured by previous methods.
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Utilization of new generation sequencing methods to elucidate cystic fibrosis-like phenotype at patients with unclear illness of molecular type.
Matějčková, Iva ; Macek, Milan (advisor) ; Holá, Dana (referee)
Cystic fibrosis (CF) is genetically conditioned, autosomal recessive disease that occurs in the European population with a prevalence of about 1:2500 - 1:1800. In this disease we observe a mutation of the CTFR gene with subsequent fault in chloride channels. Such afflicted individuals usually suffer from chronic respiratory problems, pancreatic insufficiency, high concentration of chloride ions in sweat and obstructive azoospermia. Genetic testing of CFTR gene is indicated in individuals who meet the CF clinical picture and a positive sweat test (increased concentration of chlorides in the sweat). Genetic testing of the CFTR gene is usually done by using commercial kits detecting the most common mutations of the CFTR gene in the Czech Republic. If the testing results are negative, it is further performed an MLPA method that captures the larger deletions and duplications of gene, eventually a sequencing of all exons is. Despite the well-established algorithm of the testing, some patients suffering from symptoms of CF are left without genetic findings. Thanks to development of next generation sequencing, it is possible to make the diagnosis of CF more effective and uncover the variants that were not captured by previous methods.
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Molecular genetic diagnosis of cystic fibrosis
Matějčková, Iva ; Krkavcová, Miroslava (advisor) ; Kotlas, Jaroslav (referee)
Cystic fibrosis is a very common and serious disease. It is an autosomal recessive disease with an incidence of 1 : 2500 - 1 : 4500. With this disease, CTFR gene mutations occur which are responsible for the failure of chloride channels. It is a multi-organ handicap, which primarily infects the respiratory and gastrointestinal system, but also affects the hepatobiliary system, reproductive system, and sweat glands. Cystic fibrosis can be diagnosed by a sweat test, molecular genetic diagnosis, or more rarely, using transepitel potential differences, which has not been introduced in the Czech Republic yet. Since 2009, the Czech Republic applies neonatal screening CF, which helps make a faster diagnosis to then be able to proceed to the rapid introduction of appropriate treatment. The introduction of an early treatment has significantly improved the prognosis of the patient. Currently CF patients can live on average from thirty to forty years. For the CFTR gene genetic testing is used postnatal, prenatal or preimplantation material. From these samples, DNA gets isolated and is further investigated using genetic methods, which may be based on the principle of PCR, reverse hybridization, restriction analysis, DNA sequencing or MLPA. Powered by TCPDF (www.tcpdf.org)
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