|
|
Diagnostics of genom conditioned diseases with the use of micro- and nanoparticles
Mondeková, Věra ; Prášek, Jan (referee) ; Provazník, Ivo (advisor)
The bachelor´s thesis discusses possibilities of viral genome´s detection through use of biosensors, more specifically through use of magnetic particles. The introductory part consists of brief characteristic of viruses, mentioned as originator of genom conditioned diseases, followed by chapters related to selected methods of nucleic acid´s extraction and analysis. The main part is dedicated to magnetic particles. The practical part of thesis deals with possibility of use of biosensors in specific viral pathogen´s detection, selection of biocompatible molecules suitable for magnetic particle modification and description of specific DNA sequence isolation procedure through use of magnetic particles.
|
|
|
Isolation of PCR-ready DNA from dairy products for baby nutrition
Mantlová, Jana ; Eva, Kvasničková (referee) ; Španová, Alena (advisor)
The work was focused on isolation of PCR-ready DNA and the identification of probiotic lactic acid bacteria that were isolated from five milk product for infant nutrition. DNA was isolated from crude cell-lysates of the products by magnetic P(HEMA-co-GMA) microspheres. DNAs isolated from crude cell lysates of control strains using phenol extraction method were used as positive controls. Using PCRs DNA of genera Bifidobacterium and species B. animalis, B. bifidum, B. breve, B. infantis, B. longum and Streptococcus thermophilus species were identified in products. The results obtained are consistent with the data declared by the manufacturers.
|
|
|
Selective isolation of the genus Bifidobacterium bacteria from foods
Mizerovská, Lucie ; Šárka, Havlíková (referee) ; Rittich, Bohuslav (advisor)
Probiotic lactic acid bacteria (LAB) are very often used in food procesing industry, such as milk products, cheese and fermentsd salami production in nova days. In diploma thesis were tested symbiotic food supplements from different producers. Bacterial DNA was isolated from crude cell lysates of six food suplements by magnetic particles P(HEMA-co-GMA). PCR-ready DNAs were isolated. from all products The detection of Bifidobacterium bacteria identified by PCR was in agreement with those declared by the manufacturers. Magnetic particles with immobilized antibodies against Bifidobacterium were used in the next part of thesis. These particles were used for the isolation of target cells from two products with cell identification by genus specific PCR.
|
|
|
Identification of bacteria of Lactobacillus acidophilus species in probiotic products
Sznapková, Veronika ; Trachtová, Štěpánka (referee) ; Španová, Alena (advisor)
Probiotic lactic acid bacteria (LAB) are an important part of fermented dairy products, pharmaceuticals and food supplements. At present, rapid and accurate identification of bacteria is carried out using molecular biological methods based on DNA amplification. The aim of the thesis was to identify by non-cultivation bacteria of genus Lactobacillus and bacteria of species Lactobacillus acidophilus in complex matrices at total of seven different food supplements. Total DNA was isolated from crude cell lysates using magnetic carrier P(HEMA-co-GMA). Amplificability of DNA was verified by PCR using primers specific for the domain Bacteria. In next step isolated DNA was amplified using primers specific for the genus Lactobacillus and species Lactobacillus acidophilus to demonstrate the presence of this bacterial genus and species declared by the producers. The results of bacteria identification obtained by PCR were compared with declared specification given by the producers.
|
|
|
Selective isolation of of the genus Lactobacillus bacteria from foods
Novotná, Eva ; Šárka, Havlíková (referee) ; Rittich, Bohuslav (advisor)
Probiotic lactic acid bacteria of genus Lactobacillus play an important role in the digestive tract of human. They are used in food processing and they are the part of food supplements. Lactic acid bacteria of the genus Lactobacillus can be identificated by polymerase chain reaction (PCR). Bacterial DNA was isolated from cell lysates of 4 synbiotic food suplements by magnetic particles P(HEMA-co-GMA). Isolated DNA was amplified by genus-specific and species-specific primers. Magnetic particles with immobilized antibodies against Lactobacillus bacteria were used in the next part of thesis. These particles were used for isolation target cells from products with their identification by genus specific PCR.
|
|
|
Identification of selected species of lactic acid bacteria in dairy products
Vystavělová, Růžena ; Trachtová, Štěpánka (referee) ; Rittich, Bohuslav (advisor)
Lactic acid bacteria are natural part of the human gastrointestinal tract. They are often used in food supplements and for the production of fermented dairy products. This thesis focuses on the identification of selected species of lactic acid bacteria and bifidobacteria in cheese and dairy products. Bacterial DNA was isolated by magnetic particles P(HEMA-co-GMA) from crude cell lysates from 9 products. Isolated DNA was amplified in genus-specific and species-specific polymerase chain reactions (PCR). The obtained amplicons were detected by agarose gel electrophoresis. The results of PCR were compared with those provided by the manufacturers and there has been declared a match.
|
|
|
Molecular identification of selected species of lactic acid bacteria and bifidobacteria in food additives
Riegelová, Kristýna ; Rittich, Bohuslav (referee) ; Španová, Alena (advisor)
Probiotic lactic acid bacteria and bifidobacteria are natural part of microflora of gastrointestinal tract. In the present, day they are grossly exploited in food processing industry. The aim of the work was molecular identification of bacteria of genus Lactobacillus and Bifidobacterium in complex matrices of two food additives. Total DNA was isolated from crude cell lysates by magnetic particles P(HEMA-co-GMA). Isolated DNA was amplified in genus-specific and species-specific PCRs. Amplicons were detected by agarose gel electrophoresis. Results were compared with declared specification given by producers in three different batches.
|
|
|
Isolation of DNA from probitic products using solid carriers
Bonczek, Ondřej ; Horák, Daniel (referee) ; Rittich, Bohuslav (advisor)
Microbial DNA was isolated from lysed cells of Lactobacillus genus in probiotic products. Reversible adsorption DNA on the surface of carboxyl coated nonporous poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) (P(HEMA-co-GMA)) magnetic particles and silicagel coated manganase Perovskite nanoparticles. DNA was adsorbed on the surface of the particles in the presence of 16 % poly(ethylenglycol) (PEG 6000) and 2 M sodium chloride (NaCl) concentrations. The adsorbed DNA was released from particles by low ionic strength TE buffer (pH= 8.0). The quality of isolated DNA was checked by spectrofotometric measurement and PCR amplification. DNA samples isolated using magnetic particles and phenol extraction method (control method) were PCR-ready. The DNA isolated from lysed cells of probiotic products was quantificated in real-time qPCR.
|
|
|
PCR identification of nonpathogenic bacteria strains in cheeses
Jurečková, Nela ; Doušková, Dagmar (referee) ; Španová, Alena (advisor)
Different species of genus Bifidobacterium are part of human and animal intestinal flora. These bacteria have benefit effects and therefore they are used in foods and pharmaceutical products as probiotics. Cheese is now suitable as a probiotic matrix except yoghurts and fermentated milks. This diploma thesis was focused on optimalization of DNA isolation from bacteria of genus Bifidobacterium. Magnetic microparticles (P(HEMA-co¬-GMA)) were used for DNA isolation in presence of 8% polyethyleneglycol PEG 6000 and 5 M sodium chloride. Phenol extraction weas also used as an isolation method. Isolated DNA was used for amplification in domain, genus and species specific PCRs. Optimized method was tested for detection of bacteria of genus Bifidobacterium in experimentaly prepared probiotic cheeses. These cheeses contained potential probiotic bacteria from Laktoflóra collection. Bacteria were identified into species using species specific PCR. Species Bifidobacterium animalis was identified in all samples of probiotic cheeses.
|
|
|
Specifická izolace microRNA pomocí magnetizovatelných mikročástic
Vlahová, Veronika
MicroRNAs are small non-coding RNA molecules with length of about 22 nt. These molecules participate on regulation of gene expression at the post-transcriptional level. They represent the largest group of regulators in the cell and therefore are also involved in all key processes such as proliferation, differentiation or apoptosis. Moreover, they participate in tumor transformation. These small molecules have a great potential to be diagnostic markers or assist in the treatment and prevention of diseases. This research was focused on the development of isolation method using magnetic particles with subsequent electrochemical detection of microRNA. Optimization steps were performed and then the entire method was successfully applied to real samples of HEK293 cells expressing increased levels of miR-124. The developed method proved to be sufficiently specific and applicable to the analysis of microRNA.
|
guest ::
