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Molecular Genetic Analysis in Patients Suspected of Cryptic Rearrangements.
Šolc, Roman ; Hirschfeldová, Kateřina (advisor) ; Vícha, Aleš (referee)
Such chromosomal rearrangements, which cannot be detected by using of cytogenetic banding of metaphase chromosomes, i.e. chromosomes smaller than 3 - 5 Mb, and therefore modern molecular genetic methods are used to detect them, are called "cryptic rearrangements". Their important role in human pathology is more and more significant. By using of the multiplex ligation-probe dependent amplification method (MLPA) we examined a group of 50 probands with idiopathic mental retardation. A cryptic rearrangement was found at 8 probands (16 %), at 6 of them it was demonstrably causal. Then we examined a group of 40 probands suspected of gene SHOX pathology. A cryptic rearrangement was found at 17 probands (42.5 %) and at 8 of them it was demonstrably causal. Presence of small deletion founded isolated at 7 probands was verified in a population set, but without a positive result. An analysis of mutations was made too.
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Microduplications on human chromosomes
Štolová, Lucie ; Šolc, Roman (advisor) ; Brynychová, Iva (referee)
Microduplications are small chromosomal aberrations, for whose detection it is necessary to use molecular cytogenetic methods (FISH, CGH) instead of common cytogenetic methods. Together with microdeletions, they are most often mediated by non-allelic homologous recombination during meiosis. They occur at many places in human genome and the duplications of some chromosomal regions are responsible for syndrome emergence. Some of the genes, that are included by microduplications, are dosage sensitive and they cause the pathological phenotype. As a result of development of molecular genetic methods and their usage in studies targeted on microduplications, it comes out, that presence of microduplications on the human chromosomes was undervalued, especially because of their minor clinical significance compared to microdeletions.
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Molecular Genetic Analysis in Patients Suspected of Cryptic Rearrangements.
Šolc, Roman ; Hirschfeldová, Kateřina (advisor) ; Vícha, Aleš (referee)
Such chromosomal rearrangements, which cannot be detected by using of cytogenetic banding of metaphase chromosomes, i.e. chromosomes smaller than 3 - 5 Mb, and therefore modern molecular genetic methods are used to detect them, are called "cryptic rearrangements". Their important role in human pathology is more and more significant. By using of the multiplex ligation-probe dependent amplification method (MLPA) we examined a group of 50 probands with idiopathic mental retardation. A cryptic rearrangement was found at 8 probands (16 %), at 6 of them it was demonstrably causal. Then we examined a group of 40 probands suspected of gene SHOX pathology. A cryptic rearrangement was found at 17 probands (42.5 %) and at 8 of them it was demonstrably causal. Presence of small deletion founded isolated at 7 probands was verified in a population set, but without a positive result. An analysis of mutations was made too.
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Using MLPA method for determining unbalanced changes in genome
KŘÍHOVÁ, Miroslava
Unbalanced chromosomal structural changes are connected with the presence of supernumerary particular part of chromosome, or the chromosome is absenting. MLPA is a method based on PCR principal, which amplifies MLPA probes, not the target sequences. MLPA probes hybridize to target sequences and then ligate them. Only one pair of primers is used. In the theoretical part the MLPA method is presented. Additionally, other diagnostics method of clinical genetic are mentioned (for example PCR, FISH, Array CGH). Diseases caused by unbalanced chromosomal structural changes are disscused too. Mental retardation and BRCA 1, 2 mutation were the main topic. The experimental part took place in Molecular Biology and Genetics Laboratory in the hospital in České Budějovice. I did my experiments under professional care of Mgr. Ondřej Scheinost and his colleagues. The aim of using MLPA method was to diagnose BRCA 1, 2 gene, microdeletion syndroms or subtelomeric deletions. All procedures were done according to standards. The final part of my thesis is concerned on the results interpretation and their comparation with other methods. I also thought over different approach of statistical analysis in MLPA.
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