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Diagnostics of Clostridium difficile as nosocomial infections in Nemocnice ČB a.s. using methods of molecular biology.
ŠTĚRBOVÁ, Denisa
Nosocomial infections caused by Clostridium difficile represent a substantial part of hospital-acquired infections in Czech hospitals. Intoxication by toxins produced by Clostridium difficile leads to serious damage to gastrointestinal tract and life of the patient may be in danger. The progress of intoxication could be quick, this is why reliable and time-efficient diagnostic methods are of great importance for efficient treatment of the patients. Bacterial toxins are not produced during the whole life cycle of Clostridium difficile. This is why it is better to detect bacterial DNA which is always present in the bacterial cells, not the toxins. In Nemocnice ČB a.s. (České Budějovice municipal hospital) I compared methods based on toxins detection (?hyplex? ClosTox? and ?hyplex? ClosTox 027? by BAG Health Care) with a method based on DNA detection (real-time PCR ?Xpert C. Difficile? by Cepheid). I found out the real-time PCR method is much quicker. It takes one hour to prepare the samples and to obtain analytical result for this method. Both tests based on toxin detection are much more time consuming. It takes up to 5 hours to complete them. I conclude the real-time PCR is much quicker analytical method and it allows Clostridium difficile detection during all life phases of the bacteria.
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Endoparasitosis of feathered game
BARVÍŘ, Pavel
A total of 180 samples, 119 from pheasant (Phasianus colchicus) and 61mallard (Anas platyrhynchos) were collected during two consecutive years (from 2011 to 2012). All samples were examined for the presence endoparasite using the flotation method according Sheather. In samples from pheasants were detected these types of parasites: Capillaria spp., Eimeria spp., Heterakis spp., and Trichostrongylus spp. In addition, Syngamus spp. and Cryptosporidium spp. were detected in the duck samples. Microscopical examination of aniline-carbol-methyl violet stained fecal smears revealed to Cryptosporidium spp. 12 positive samples originating from Anas platyrhynchos. DNA was extracted from Cryptosporidium positive samples and all microscopically negative samples. Nested PCR was performed to amplify the partial SSU rRNA gene of Cryptosporidium. The sequence analyses of PCR-positive specimens identified 10 samples as Cryptosporidium avian genotype III and 4 samples as C. muris. No clinical signs were detected in any Cryptosporidium positive animals. This is the first report of Cryptosporidium avian genotype III in the Czech Republic and also in Anas platyrhynchos.
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Congenital disorders by pigs
MUSILOVÁ, Dagmar
This diploma thesis follows the bachelor thesis and it?s task is to processing analysis of the occurrence of PSE meat in a population of the Czech white breed as a pigs stress syndrome impact. General problem of congenital diseases is processed in literature review. Further, this work deals with pigs stress syndrome and stress, which causes the meat defects as an external factor. Molecular - genetic methods, which are used in the practical part, are described as well. The research focused on genotyping of loci selected panel of samples, to determine the frequency of alleles and genotypes of selected locus and the statistical relationship between genotype and expression of PSE meat. Results were statistically processed and evaluated at the end.
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Molecular and biochemical characteristics of genetically modified barley plants
KOCKOVÁ, Lucie
Modern agriculture often employs broad-spectrum herbicides combined with herbicide-resistant crops. Usually, this is achieved by altering the crop genom (genetic modification) by inserting of a specific gene coding for resistance to specific herbicide or group of herbicides. Apllying such herbicide thus results in minor crop damage. The resistence of crops against broad-spectrum herbicides depends on the genes, which were inserted into the genom. The gene bar is often used as a resistence -providing element. It mediates resistance against a broad spectrum of herbicides, such as glufosinate and Bialahops. For the selection of transformants and preliminary assessment of the target transgen expression level a suitable marker gene is simultaneously inserted. For this purpose, gene for bacterial ??glucuronidase (GUS) is often used. It is considered to be one of the most frequently used reporter systems for the assessment of transgen expression and also allows to analyze the expression on the tissue, cell and whole cell organelles levels. In this diploma thesis the PCR method and histochemical detection of the enzyme glucuronidase presence were used to detect and evaluate transgenic plants of cv. Golden Promise spring barley, modified by genes bar and gus. The presence of gus gene was determined in different parts of plants.
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Methods of DNA extraction from the fresh papaya fruit and from the candied
Ovesná, Jaroslava ; Hodek, Jan
Aim the work is to provide a working procedure for extraction of amplifiable DNA form papaya fruits (Carica papaya), which are a poor source of DNA. Methods are optimised for DNA extraction from papaya fruits and stones and from candied papaya- these both were predicted as suitable commodities for an eventually screening of non-approved GM papaya in the market of Czech Republic. The methodology was prepared on basis of demand of reference laboratories of public service, which are involved in GMO analyses and also as a flow-work of Methodics for Detection of GM papaya 55-1 and 63-1 (Hodek, Ovesná, Pavlátová 2008) and scientific publication Detection of transgenic papaya lines: extraction protocol optimisation and verification of DNA quality by PCR assay (Ovesná, Hodek 2009).
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Genotyping of \kur{Giardia intestinalis} isolates
ŠRÁMOVÁ, Eliška
The aim of this work was assemble isolates of Giardia intestinalis from humans and other mammals. Stools samples were examined for presence of cysts by concentration settling method. Consequently sequencing of 532 bp parts of the TPI gene after previous amplification by the nested PCR was performed. In vitro cultures of selected isolates were established using experimental model hosts, gerbils.
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