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Analysis of gonosomal mosaic using FISH method
ŠUTKOVÁ, Jana
In my bachelor thesis I dealt with the analysis of gonosomal mosaics. Mosaicism can be described as a condition in which two or more distinct cell lines originating from a single zygote are found in one organism. In women, the most common mosaic forms are Turner syndrome and triple X syndrome or superfemale. In the male population, Klinefelter syndrome and XYY syndrome (supermale) are most commonly detected. Mosaicism can be investigated by fluorescence in situ hybridization. In the theoretical part of my bachelor's thesis I focused on congenital chromosomal mosaicism, especially on its variants such as Klinefelter syndrome, Turner syndrome, triple X and XYY syndrome. Furthermore, in this part I dealt with fluorescence in situ hybridization. I described fluorescence per se and then its application in clinical practice, where several types of fluorescence in situ hybridization are used. In the practical part I examined the number of gonosomal chromosomes using fluorescence in situ hybridization. I prepared this part in the laboratory of the Faculty of Health and Social Sciences of the University of South Bohemia in České Budějovice. I examined 40 anonymized samples from 20 probands. Capillary blood samples and buccal swabs were taken from each of them. After hybridization of samples with centromeric probe for X and Y chromosomes microscopy was performed. In the last part of this bachelor thesis I dealt with the processing of the results obtained by fluorescence microscopy. Thus, a total of 6 cases of mosaicism were detected in women (the percentage of mosaic cells was above the admissible limit). In males there were a total of 2 cases of increased percentage of mosaic cells.
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The increased diagnostic efficiency of QF-PCR for aneuploidy of amniotic fluid
Sedláková, Zdeňka ; Macek, Milan (advisor) ; Daňková, Pavlína (referee)
Quantitative fluorescence polymerase chain reaction (QF-PCR) is a molecular genetic method based on the amplification of microsatellites (Short tandem repeats, STR) and measurement of the peak heights of amplicons in the electropherogram. Currently, the QF-PCR deemed reliable, fast, and inexpensive method that is gradually replacing conventional cytogenetic analysis of aneuploidy (examination of long-term cultures of amniotic fluid). However, in certain cases it is impossible to determine the parental origin and meiotic aneuploidy by QF-PCR. The aim of this work was to verify the new dinucleotide STR markers on chromozomes 13, 16, 18, 21, and 22 and further increase the diagnostic efficiency of QF-PCR retaining other STR markers on chromozome 15, 16, 22 and to determine the population and the analytical characteristics of these markers. For all dinucleotide STR markers stutter occurred in high frequency and therefore there were found not to be suitable for routine diagnostics. STR markers for chromozomes 15, 16 and 22 were tested on 100 patients. We selected four informative markers for both chromozome 16 and 22, and three markers for chromozome 15. Thus, I expanded set of diagnostic STR markers in this thesis.
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The increased diagnostic efficiency of QF-PCR for aneuploidy of amniotic fluid
Sedláková, Zdeňka ; Macek, Milan (advisor) ; Daňková, Pavlína (referee)
Quantitative fluorescence polymerase chain reaction (QF-PCR) is a molecular genetic method based on the amplification of microsatellites (Short tandem repeats, STR) and measurement of the peak heights of amplicons in the electropherogram. Currently, the QF-PCR deemed reliable, fast, and inexpensive method that is gradually replacing conventional cytogenetic analysis of aneuploidy (examination of long-term cultures of amniotic fluid). However, in certain cases it is impossible to determine the parental origin and meiotic aneuploidy by QF-PCR. The aim of this work was to verify the new dinucleotide STR markers on chromozomes 13, 16, 18, 21, and 22 and further increase the diagnostic efficiency of QF-PCR retaining other STR markers on chromozome 15, 16, 22 and to determine the population and the analytical characteristics of these markers. For all dinucleotide STR markers stutter occurred in high frequency and therefore there were found not to be suitable for routine diagnostics. STR markers for chromozomes 15, 16 and 22 were tested on 100 patients. We selected four informative markers for both chromozome 16 and 22, and three markers for chromozome 15. Thus, I expanded set of diagnostic STR markers in this thesis.
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The occurrence of chromosomal aberrations in the Czech Republic
LANDOVÁ, Ivana
The objektive of this bachelor's thesis was to describe the occurrence of chromosomal aberrations in the Czech Republic and to verify whether the literary data agree with the current occurrence rates. This thesis consist of 2 parts: The theoretical part of the thesis describe phenotypic symptoms and signs of chromosomal aberrations, their origin, causes and occurence rates in the population as reported in the literature. In the practical part, I tried to verify these occurrence rates of the aberrations based on results of investigations (both prenatal and postnatal) provided by the following laboratories: Genetika Plzeň s.r.o., Institute of Reproductive Medicine and Genetics, Karlovy Vary, and Institute of Medical Genetics, University Hospital, Pzeň. I have also used data obtained from the Czech Institute of Medical Information and Statistics (ÚZIS) as well as data found in various Internet sites. The purpose of this research was to provide graphics demonstrating the results, unify various types of results and try to find possible interrelations. It is obvious from the results that there has been a shift in the age group of maximum fertility from ages 20 ? 24 in 1995 to 30 ? 34 years in 2008. The mean age of mothers whose foetuses were diagnosed with Down syndrome in individual years ranged in the interval from 27 to 37.5 years. There were no differences in the representation of Down syndrome in individual years, and also the representations found in individual workplaces did not show any significant differences, ranging from 0.19 to 1.52%. The mean occurrence rate of Down syndrome as calculated from the results published by ÚZIS from the period 1994 ? 2007 was 1 for 1,700 live births. I succeeded in realizing all the targets of my work. Both prenatal and postnatal occurrence rates of aberrations in the workplaces of interest varied for individual years or showed only minor signs of increasing or decreasing rates. Monitoring of larger number of workplaces appears warranted to confirm my hypothesis.
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