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Comparison of methods for determination of Neisseria meningitidis in Nemocnice ČB a.s.
SKOŘEPOVÁ, Lucie
Molecular biology methods are of great importance for routine diagnostics of infections diseases. In this work two such methods were compared for the case of diagnostics of Neisseria meningitides in Nemocnice ČB a.s. Nemocnice ČB a.s. is a municipal hospital owned by South Bohemia Region. I hypothesised that real-time polymerase chain reaction provides lower analysis time compared to nested polymerase chain reaction. These two methods are routinely used in Central Laboratories of the hospital and it is feasible to use both of them for the diagnostics of Neisseria meningitides. During the investigation I have found another method useful for the diagnostics of this pathogen. Latex agglutination test is an immunological reaction used to detect antibodies which appear as an immune response to infection. In spite of very quick output, the immunological origin of the latex agglutination test brings serious limitations: any suppression of the immune response or degradation of the antibodies leads to false negative results of the test. Lower reliability of the latex agglutination test compared to various polymerase chain reactions based on the detection of bacterial DNA strengthen my devotion to compare the two methodologies of reliable polymerase chain reaction and led to a decision not to include the latex agglutination test into the original research plane. In good agreement with the original expectation, practical evaluation of tests based on real-time polymerase chain reaction and on nested polymerase chain reaction displayed significant differences in the analysis time (3 h 40 min versus 7 h 30 min respectively). Another advantage of the test based on real-time polymerase chain reaction is its higher sensitivity of one order of magnitude more than in the test based on nested polymerase chain.
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Congenital disorders by cattle
KOSOBUDOVÁ, Hana
In the framework of this thesis was performed genotyping of 46 specimens of the breed Czech red cattle from the University farm in Czech Budejovice, which monitored the incidence of autosomal recessive genetic disorders, specifically bovine citrullinemia (BC) in exon 5 and deficiency of blood coagulation factor XI (FXI) in exon 9 and 12. Genotyping for BC was done using PCR/RFLP methods and for the disorder FXI in both exons genotypes were determined on the basis of different length of fragments using PCR technology and horizontal agarose electrophoresis. The presence of mutant allele was detected only in the locus for BC and that is in 7 heterozygous carriers, who produced three bands with a length of 185 bp fragments, 103 bp and 82 bp. The frequency of mutant allele and the frequency of heterozygous carriers to 7.6% and 15.2%. Results of the study show that the presence of mutant allele for BC in our tested panel of animals is relatively high. In the future it will be necessary to adopt measures that will lead to the elimination of this allele. Otherwise, its further dissemination would have a negative impact on the health of the population and there might occur complications in the regeneration of Czech red cattle, which is one of our farm animal genetic resources. The literature review deals with the problems of congenital disorders and discusses the importance of health heredity and understanding of the genomic information of cattle.
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Evaluation of SDS-PAGE tuber protein profiles for their potential use in potato cultivar characterization
PEŠINA, Jiří
The use of various types of gel electrophoresis for characterization and identifying plant varieties is well estabilished. This diploma work deals with the possibility characterize potato cultivar (Solanum tuberosum L.) by the help of electrophoretic technique SDS {--} PAGE tuber protein profiles. Soluble potato tuber proteins can be classified according to their molecular weight (kDa) into three groups: First, the major tuber protein patatin (37 {--} 44 kDa); second, a group constituted of protease inhibitors (6 {--} 24 kDa); and third group containing all other proteins. First two group manifest genetic variability, which is necessary evalute for its potential using of potato cultivar characterization.
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Fylogeography and genetics variability of \kur{Diuraphis noxia} (\kur{Aphididae})
PAŠÍKOVSKÝ, Jiří
The aim of this work was a research of the genetic variability of natural populations of Russian wheat aphid Diuraphis noxia (Aphididae) by means of microsatellite markers and markers for EPIC-PCR. First goal was to introduce the methods and optimise them for Diuraphis noxia. In the follow-up pilot study, specimens from 47 lines representing 12 populations from all over the world were analysed. Having used microsatellite markers, I proved expected variability among individual populations and within them. The highest genetic variability was detected between Chile and Algeria using markers for cytochrome C in EPIC-PCR. These findings can be used for further studies of the genetic variability of the aphid Diuraphis noxia.
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