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Methods of molecular diagnostics of parasitic helminths
Dejmková, Tereza ; Leontovyč, Roman (advisor) ; Škorpíková, Lucie (referee)
Helminthiases represent a significant health risk, therefore the use of effective and rapid diagnostic methods is essential, including molecular methods, which are summarized in presented thesis. The biology of important human helminth pathogens is summarized in the introductory part. The DNA isolation, visualisation and molecular methods are described in the following parts. The section about molecular methods is divided into two parts. The first part is focused on PCR and its modifications, the second part describes methods of isothermal DNA amplification, which represent more practical and effective alternative to PCR. The advantages, disadvantages of all methods and examples of helminthiases to which they have been applied are also discussed. The final part is about the impact of the COVID-19 pandemic on the development of molecular diagnostic methods, during which were previously established protocols modified or new molecular methods were applied to increase the efficiency. The presented thesis brings an overview of molecular methods that were used to this day for diagnosis of helminthiases. At the same time the thesis discusses the impact of the COVID-19 pandemic on the development of molecular diagnostics and shows the possible future development of molecular diagnostic methods in the field of...
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Optimization of a loop-mediated isothermal amplification method for a detection of Haemophilus influenzae infection.
Zadáková, Kateřina ; Jeřábek, Petr (advisor) ; Svášková, Dagmar (referee)
Haemophilus influenzae is a gramnegative, rod-shaped and predominantly human pathogen. Infection caused by this microorganism is especially dangerous for children under five years of age, who are mainly at risk of pneumonia and meningitis.Althought vaccination is available against its most dangerous type b, it still causes problems, especially in developing countries. Due to the insufficient vaccination, strains that are more resistant to the antibiotics are more spreading in these countries. Equipment requirement of current detection options for H. influenzae infection(e.g. polymerase chain reaction) causes, that reliable detectionis a problem in poor countries. Loop-mediated isothermal amplification could be a cheap and reliable method. The aim of this bachelor thesis was the design of sets of primers and their applicationinthe LAMP reaction. The designed primers targetedthe ompP6 gene, which was alreadythe target of LAMP reaction in some studies and is also a frequent target in PCR detection.As a part of the optimization, different volumes of the reaction mixtures and multiple detection options were tested -colorimetric detectionusinganacid-base indicator and fluorescence detection using different dyes monitored in real time. Key words: Haemophilus influenzae, loop-mediated isothermal...
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Světelný objekt inspirovaný japonskou tradicí
Kogure, Haruka
The aim of this thesis was to examine traditional Japanese lamps and the origami technique. Also, to learn about modern trends in forms of illumination and compare their design among Japan and other countries. From the gathered knowledge a new illumination object inspired by Japanese tradition was designed.
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Small ruminant lentiviruses - analysis of disease prevalence and distribution of SRLV genotypes in sheep and goat farms in the Czech Republic
VERNEROVÁ, Kateřina
Maedi-visna in sheep, and arthritis and encephalitis in goats, are globally widespread and progressive inflammatory diseases caused by retroviruses belonging to the small ruminant lentivirus (SRLV) group. They cause lifelong infections in affected animals, characterised by slow progression to overt disease and are always fatal. The aim of this study was to determine the prevalence of SRLV disease in the Czech Republic, to map the genotypic representation of SRLV using phylogenetic analysis, and to analyse TMEM154 as a selected candidate marker of SRLV resistance in sheep and goats. A total of 3 410 sheep and goat blood samples were collected from 21 flocks. The seroprevalence of maedi-visna in sheep was found to be 19.9% (556/2801), and the seroprevalence of arthritis and encephalitis in goats was 14.1% (86/609). All seropositive animals were tested by nested polymerase chain reaction (nPCR) for the presence of proviral DNA. Phylogenetic analysis identified the SRLV genotype in 77 sequences, of which 60 sheep and goat samples were genotype A and 17 sheep samples were genotype B. While all genotype B sequences were classified as subtype B2, the group of genotype A isolates showed higher variability and were related to subtypes A2 and A3. In addition, 40 seropositive and 50 seronegative sheep and goat samples were randomly selected to design a methodology for LAMP (loop-mediated isothermal amplification) diagnosis of SRLV in sheep and goats. Seronegativity was clearly confirmed by the LAMP method in all samples, and seropositivity was confirmed in 31 out of 40 cases in both sheep and goats. 605 sheep and 60 goat samples were randomly selected for TMEM154 genotyping. Most seropositive animals were heterozygous EK (61%), 58% were homozygous EE, and 45% were homozygous KK. In sheep, all 3 genotypes were identified, while all goats were homozygous EE.
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Detection MDR1 mutation in shepherd dog breeds by loop mediated isothermal amplification and restriction analysis
VYHLÍDALOVÁ, Nikola
Multiple drug resistance is caused by MDR1 gene mutation. It is a common genetic disease of Shepherd Dogs from which Collie breed is the most affected. Described mutation is caused by deletion of four nucleotides, which leads to a frameshift and premature termination of synthesis of the P-glycoprotein, one of the important transport proteins of some drugs. Synthesized non-functional P-glycoprotein is unable to eliminate drugs from the brain, which leads to a neurotoxic problems in affected dogs. The theoretical part describes structure and function of P-glycoprotein and its inhibition caused by MDR1 coding gene mutation. The most commonly affected breeds are subjected to analysis as well as the problem of drugs for dogs with this mutation. Review also includes a description and principle of methods used for detecting mutation of MDR1 gene. The aim of this thesis was to optimize and screen DNA samples of Shepherd Dogs for the MDR1 gene mutation by LAMP and PCR-RFLP as well as by mutual comparison of both methods. The empirical part presents the results of the study. The studied population consisted about 140 samples of Shepherd Dogs DNA. Our results confirmed the highest incidence of recessive homozygotes in the tested population of Smooth Collie (25 %). Moreover, the number of heterozygous was also the highest in this breed (50 %). However, the number of heterozygous dogs in the tested population of German Shepherd was surprisingly comparable with population of Smooth Collie, because it reached 49 %. Equal number of the German Shepherd was dominantly homozygous. Whereas, the highest number of healthy dogs was found in the tested population of Border Collie breed (65 %).
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Production of ceiling light
Kvasnička, Jan ; Řiháček, Jan (referee) ; Císařová, Michaela (advisor)
The manufactured component is the inner part of the ceiling lamp. It is made of 17 240 steel in a series of 100 000 pieces/year. A combination of tool cutting and deep drawing without thinning the wall was chosen from the possible production technologies. The diameter of the blank from which the resulting shape is pulled out was calculated to be 400 mm. It will be cut from an unwound coil of sheet metal, the inner diameter of which is 400 mm and the outer 1000 mm. A drawing tool was designed for forming and it's functional parts were made of 19 436 steel. The tool is placed in hydraulic three-acting press PO 160/100 from company Dieffenbacher. The press is part of a line which includes an unwinding device, a straightening device and a cutting tool.
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Design of Outdoor Wireless Speaker
Kráčmarová, Gabriela ; Haltof, Vladimír (referee) ; Sovják, Richard (advisor)
Thebachelor thesis dealswiththe design ofanoutdoorwirelessspeaker. Theaimwas to design a functionalfriendly design thatwillwithstandadverseeffects in nature. Theproductisdesignedwithergonomics, weight (up to 750 g), additionalfeatures (lighting and FM) and technology in mind. Emphasiswasplaced on comfortabletransmission, innovativeappearance, pleasant and easyhandling, enriched by additionallightingfunctions (decorative, warning, stroboscopic, replacingthe lamp and lantern), Power Bank and listening to radiostationsthanks to FM. It is a speakerwithadaptedpartsfor a suitablegrip (handles), abounding in surroundsound and with a fullyroundedcover made ofdurablematerial. Theenduranceisprotected by a li-pol 8,000 mAhbatterywithrechargingfrom USB C. The design istargetedattheactive part ofthepopulation, whichcan use it in thefield (IP67 protection).
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Detection of Ralstonia solanacearum bacteria pathogenic for tomato, pepper and potato by loop-mediated isothermal amplification
STRNAD, Adam
For rapid detection of quarantine bacteria Ralstonia solancearum pathogenic for tomato, potato and papper, the LAMP method was used. Based on the analysis of the fliC gene sequence, a set of LAMP primers was designed to be specific for Ralstonia solanacearum. Subsequently, the method was optimized to detect only Ralstonia solanacearum and to distinguish it from the other phytopathogenic bacteria which infect potato, tomato and pepper. LAMP can detect bacteria from very small amounts of DNA in less than 30 minutes in real time. The method is capable of amplifing DNA at a concentration of 0.00001 ng/?l. However, a higher DNA concentration is more suitable for more specific detection.
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Electronic ballast for sodium lamp
Eliáš, Jaroslav ; Vorel, Pavel (referee) ; Huták, Petr (advisor)
The aim is to get acquainted with the way the management and regulation of sodium lamps. This knowledge is then applied to design your own gear. The introductory section describes the basic characteristics of discharge light sources. Their behavior was verified by measuring and findings were used to create diagrams, mainly the control circuit. Theoretical design is developed by individual functional parts of the inverter.
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