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Detection of mutation causing plycystic kidney disease
SLÁDKOVÁ, Nikola
In my bachelor's thesis, I dealt with the detection of the mutation in the PKD2 gene, which causes polycystic kidney disease. The disease is interesting because it is not only caused by a single specific mutation that affects a particular gene, but the disease is highly variable and can be caused by several mutations located on different chromosomes and even at different sites in a given exon. The subject of my research was a family of four. Based on the results of using clinical diagnostics, one member of the family was confirmed to have polycystic kidney disease. The aim of this thesis was to detect the possible PKD2 mutation in the family members, and thus to confirm the presence of polycystic kidney disease. The next aim was the optimization of the designed primers for the PCR reaction. In the theoretical section of this thesis, I focused on the issue of polycystic kidney disease, classification of polycystic kidney disease based on the inheritance type, and the protein products of the PKD2 and PKD1 genes. I also mentioned the anatomy of the kidneys, their function, and also the structure of a nephron. I also dealt with the health problems that accompany the disease, as well as its diagnostics and subsequent treatment. In the practical section of this thesis, I described the process of the laboratory diagnostics used to search for a mutation in the PKD2 gene, specifically in its first five exons. The diagnostics consisted of the DNA isolation, PCR reaction, gel electrophoresis, and subsequent Sanger sequencing. However, due to limited laboratory capacity, I did not personally perform the Sanger sequencing. The samples prepared for sequencing were sent to the SEQme company. In the end, I examined the electrophoreograms and the results from the obtained sequences. I had a total of 20 sequenced samples for the evaluation, always 4 samples for each exon. I was not able to prove the possible mutation in any of them. The remaining exons would also need to be examined for a complete diagnostics of the disease.
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Genetically determined progression factors of selected chronic nephropathies
Obeidová, Lena ; Reiterová, Jana (advisor) ; Skálová, Sylva (referee) ; Vodička, Radek (referee)
Polycystic kidney disease is a severe genetic disease occurring in both adult and pediatric patients. The basic characteristic of this disease is the development and progressive enlargement of renal cysts gradually replacing functional kidney tissue. This leads to renal failure in many patients. However, renal cysts may also occur in a number of other diseases, including multisystem syndromes. This complicates differential diagnosis in some patients. In our study, we first focused on the diagnosis and characterization of genotypic-phenotypic relationships in patients with polycystic disease arising in childhood, later we extended our study to adult patients and patients with unclear clinical diagnosis. At the same time, we expanded the portfolio of analyzed disorders to a number of diseases in which the phenotype of polycystic kidneys may occur, and noncystic diseases as well. During our project, massive parallel sequencing was used to analyze 149 patients - 128 with cystic and 21 with noncystic clinically diagnosed nephropathies. At the same time, the findings were verified by Sanger sequencing in 176 relatives of our probands. Mutation detection reached 59% in cystic patients, and 43% in non-cystic patients, respectively. In many patients, molecular genetic analysis revealed a different etiology...
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Genetically determined progression factors of selected chronic nephropathies
Obeidová, Lena ; Reiterová, Jana (advisor) ; Skálová, Sylva (referee) ; Vodička, Radek (referee)
Polycystic kidney disease is a severe genetic disease occurring in both adult and pediatric patients. The basic characteristic of this disease is the development and progressive enlargement of renal cysts gradually replacing functional kidney tissue. This leads to renal failure in many patients. However, renal cysts may also occur in a number of other diseases, including multisystem syndromes. This complicates differential diagnosis in some patients. In our study, we first focused on the diagnosis and characterization of genotypic-phenotypic relationships in patients with polycystic disease arising in childhood, later we extended our study to adult patients and patients with unclear clinical diagnosis. At the same time, we expanded the portfolio of analyzed disorders to a number of diseases in which the phenotype of polycystic kidneys may occur, and noncystic diseases as well. During our project, massive parallel sequencing was used to analyze 149 patients - 128 with cystic and 21 with noncystic clinically diagnosed nephropathies. At the same time, the findings were verified by Sanger sequencing in 176 relatives of our probands. Mutation detection reached 59% in cystic patients, and 43% in non-cystic patients, respectively. In many patients, molecular genetic analysis revealed a different etiology...
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