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Determination of the concentration of alpha-1-antitrypsin in the stool by immunoassay method
Plačková, Marie ; Kocna, Petr (advisor) ; Průša, Richard (referee)
Determination of the concentration of α1 - antitrypsin in the stool is a diagnostic indicator of inflammatory diseases of the small and the large intestine, especially malabsorption syndrome. α1 - antitrypsin belongs to the family of plasma proteins with antiproteinase effect. α1 - antitrypsin is synthesized in liver, in small amount in macrophage and is a protease inhibitor of serine proteases sercected from neutrophils. α1 - antitrypsin is acute phase protein. Higher α1 - antitrypsin values are in early phase of inflammation associated with raised CRP and other pozitive acute phase proteins. Fecal α1 - antitrypsin clearance is a sensitive and specific marker of protein loss. For α1 - antitrypsin determination in stool samples ELISA method can be used. ELISA is noncompetetive immunoassay used to detect presence of antibody or an antigen in a sample. The aim of this work was to compare two ELISA sets (Immundiagnostik and Ridascreen) used for determination α1 - antitrypsin in the stool. Then examine stability of α1 - antitrypsin in the stool and in extract prepared from stool in various storing conditions temperature and time. After this establish this method as routine in laboratory. 20 patient stool samples were examined to compare ELISA sets. Samples were suggested to be α1 - antitrypsin...
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Individual ....
Tomeček, David ; Kocna, Petr (advisor) ; Průša, Richard (referee)
13 C-MTG breath tests are used for exocrine pancreas diagnostics. It is a quite expansive test and its usage has never been tested in clinical practice in long time horizon. This study is engaged in pre-analytical phase and contains a group of 4 people who were tested for 6 months and this is the first study ever that has been proceeded in a long time horizon. The half-year profile of these 4 people showed that some of those results are spread of tens percents against its average. The test of CO2 stability showed that the value of CO2 is independent on the temperature which the breath bag is stored in. Its concentration was same for the time of 7 days. Now we can say that the breath bags are insensitive in case of transporting and storage. The question is, why the value of delta 13 C/12 C changed. The results of this study point on fact that values of cumulative dosage recovery of these 4 people did change.
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Immunoflulorescent detection of endomysial antibodies (EmA) in the coeliac diagnosis
Martikán, Petr ; Kocna, Petr (advisor) ; Průša, Richard (referee)
The aim of this thesis was comparing of a several immunofluorescence methods and substrates for detection endomysial antibodies (EmA). The common method (Anti-Endomysium antibodies AEA) is used by UKLBD in VFN and by the some samples gives ambigious results owing to another autoantibodies (e.g. ANA, AMA, ASMA). The common method was compared with another sets (IFA-EmA-IgA - AEMAk 03a-10, ImmuGlo anti Endomysial Antipody EMA IFA Kit). Another aim of the work was how to work with this method and be able to evaluate samples by the fluorescence microscope. (...) This thesis brought me a lot of valuable experiences in laboratory and also teached me how to evaluate endomysium antibodies by indirect immunofluorescence method.
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Laboratory functional diagnostics of malabsorption syndromes with the focus on lactose intolerance
Pluhařová, Apolena ; Kocna, Petr (advisor) ; Kohout, Pavel (referee)
This diploma thesis deals with laboratory functional diagnostics of malabsorption syndromes. Especially the functional diagnosis of lactose intolerance. The golden standard for the functional diagnosis of lactose intolerance is the breath test. The aim of the study was to determine the prevalence of lactose intolerance (LI) using a lactose breath test when consuming a dose of 20 g of lactose in patients with gastrointestinal symptoms and in volunteers who do not show gastrointestinal problems. Part of the research was also to examine the symptoms that led the individual to see a doctor and undergo a lactose breath test. The method consisted in the evaluation of the results of breath tests, which were performed on patients and volunteers in the gastroenterological laboratory of the 1st Medical Faculty of Charles University and the General Hospital. A questionnaire was given to everyone to evaluate the symptoms of LI and the phenomena leading to malabsorption. Lactose intolerance was confirmed in 92 % of patients and 79 % of volunteers. The most reported symptoms in the patient sample were typical gastrointestinal LI-related problems: gastric and intestinal problems after milk consumption (85 % of patients), gastric and intestinal problems after consumption of dairy products (85 % of patients),...
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Determination of the concentration of alpha-1-antitrypsin in the stool by immunoassay method
Plačková, Marie ; Kocna, Petr (advisor) ; Průša, Richard (referee)
Determination of the concentration of α1 - antitrypsin in the stool is a diagnostic indicator of inflammatory diseases of the small and the large intestine, especially malabsorption syndrome. α1 - antitrypsin belongs to the family of plasma proteins with antiproteinase effect. α1 - antitrypsin is synthesized in liver, in small amount in macrophage and is a protease inhibitor of serine proteases sercected from neutrophils. α1 - antitrypsin is acute phase protein. Higher α1 - antitrypsin values are in early phase of inflammation associated with raised CRP and other pozitive acute phase proteins. Fecal α1 - antitrypsin clearance is a sensitive and specific marker of protein loss. For α1 - antitrypsin determination in stool samples ELISA method can be used. ELISA is noncompetetive immunoassay used to detect presence of antibody or an antigen in a sample. The aim of this work was to compare two ELISA sets (Immundiagnostik and Ridascreen) used for determination α1 - antitrypsin in the stool. Then examine stability of α1 - antitrypsin in the stool and in extract prepared from stool in various storing conditions temperature and time. After this establish this method as routine in laboratory. 20 patient stool samples were examined to compare ELISA sets. Samples were suggested to be α1 - antitrypsin...
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Determination......
Mrňák, David ; Kocna, Petr (advisor) ; Průša, Richard (referee)
The main interest of this work is in determination of pancreatic elastase 1 in the stool. The determination of the value of the pancreatic elastase 1 in the stool is important for specify of the function of the exocrine pancreas and it have high specificity and sensitivity. It is used for both long observation of patients with chronically insufficiency of the pancreas, also as a screening test for pancreatic illness and in differential diagnoses of malabsorbent syndrome. We can determine values of the pancreatic elastase 1 in the stool with using the ELISA method. ELISA means enzyme linked immunoassay and it principle is based on sandwich reaction between two monoclonal antigens with high specifity against human pancreatic elastase 1. The main objective is to determine the degree of variability in the values of the pancreatic elastase 1 in group of healthy subjects. The study will take place on four individuals and their samples will be investigated with the ELISA method in monthly intervals for six months. ELISA method was processed by a set from firm ScheBo®. After the completition of the test it is clear, that there is a high fluctuation of the values of the pancreatic elastase 1 in this group. Values of the difference between each individual measurement exceeded the limit of 70 percent....
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