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Analytical parameters for the determination of anti-HCV and HCV antigen
SCHOVANCOVÁ, Zuzana
Hepatitis C is a global health problem that affects millions of people and is a major cause of liver disease, including cirrhosis and hepatocellular carcinoma. Due to the asymptomatic course of many infections, effective laboratory diagnosis is needed. The aim of this bachelor thesis was to perform a comparative analysis of two diagnostic methods for the detection of anti-HCV and HCV antigen in serum and plasma samples from patients and to assess their diagnostic accuracy by comparison with quantitative PCR and specific immunoblotting. The methods used in the study were chemiluminescent microparticle immunoassay (CMIA) on the Architect i2000SR and electrochemiluminescent immunoassay (ECLIA) on the Cobas Pure. The scientific contribution of this thesis lies in the evaluation and description of the diagnostic performance and limitations of both test methods in laboratory practice, which may contribute to a more accurate diagnosis of hepatitis C and possible improvement of diagnostic procedures, which has important implications for clinical practice, especially in relation to the selection of the appropriate diagnostic method depending on the clinical context of the patient.
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Risk behaviour of people who inject drugs following treatment of hepatits C
Sedláková, Tereza ; Mravčík, Viktor (advisor) ; Janíková, Barbara (referee)
Background: In Czech Republic, 75-100 % of people newly diagnosed with hepatitis C type (VHC) are people who inject drugs (PWID). Currently, available treatment with direct-acting antiviral medicines without significant adverse side effects is fully covered by insurance. However, there is a risk of reinfection and insurance companies usually don't reimburse further treatment anymore. Aim: The aim of this study is to map and describe the development of risk behaviours in clients of low-threshold treatment programmes of the SEMIRAMIS and Laxus organisations (LSA partners) who have received treatment for hepatitis C. The research focuses on the circumstances of treatment, particularly barriers and supports to entry and the actual course of treatment. The sub-objectives are: to map the socio-economic situation of clients, to approximate patterns of use and to describe contacts with other services. In addition, to reveal the links between these factors and any changes in risk behaviour. Methods: Research was completed by metod of cross-sectional questionnaire surveys. The research sample consisted of 32 clients of LSA partners' low-threshold programs who had received at least one VHC treatment. The risk behaviours were monitored among following domains: a) injection use in general, b) unprotected...
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Comparison confirmation ELISA to recomLine test for the determination of hepatitis C
BAČKOVÁ, Marcela
My bachelor thesis deals with hepatitis C disease and its laboratory diagnosis, especially confirmatory tests. The theoretical part focuses on the disease in general, describes the hepatitis C virus that causes the disease. It also discusses the acute and chronic phases, virus transmission and treatment options. In the practical part I describe the examination of anti-HCV screening antibodies and how to proceed in case of a positive finding. The main task of my practical part is to compare two confirmatory tests - ELISA and immunoblot recomLine. At the end of the work I evaluate both tests and compare the final results.
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Efficacy and treatment adherence to therapy of chronic HCV infection in people who inject drugs
Jandová, Zuzana ; Gabrhelík, Roman (advisor) ; Věchetová, Sonja (referee) ; Fraňková, Soňa (referee)
Backgrounds: Intravenous drug use represents the main route of transmission of hepatitis C virus infection (HCV). IVDU represent approximately 75-100% of newly diagnosed cases of HCV. IVDU have traditionally been considered as difficult-to-treat, owing to a low treatment adherence. The presumed low adherence to care in IVDU represent nowadays the barrier to therapy with direct-acting antivirals (DAA). Aims: The aim of the thesis was to assess efficacy of therapy of HCV with DAA and adherence to therapy in patients with history of IVDU. Comparing the results with the group of patients without IVDU, we would support the fact that the treatment efficacy in the IVDU group is high and that the utilisation of financial resources is effective. Methods: All consecutive patients who started DAA anti-HCV therapy at the Department of Hepatogastroenterology of Institute for Clinical and Experimental Medicine from 1st January 2017 to 6th August 2018 were included. The patients were divided into two groups: individuals with history of IVDU (IVDU, N = 101) and control group (Controls, N = 177), without IVDU in the past. The patients' data were obtained from patients medical charts during treatment and follow-up. Results: IVDU group achieved an SVR12 of 98% (vs. 98% in controls, N. S.). Two IVDU patients who were...
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Psychosocial Problems in Patients with Chronic Hepatitis C and Drug Addiction in their History
JAROŠOVÁ, Eva
This diploma thesis deals with the psychosocial problems of patients treated for chronic hepatitis C. The aim of the thesis was to map the problems arising during this treatment and to divide them into biological, psychological, and social problems and problems leading to a change in the patient's quality of life. The disease is also assessed in relation to risky behaviour - a history of substance dependence. The work should also show how the individual components of the multidisciplinary team are involved in intervention during treatment. In the first part of the thesis, I describe the theoretical issues of hepatitis C as a consequence of intravenous drug dependence, and I deal with quality of life, the multidisciplinary team and psychosocial care with the help of professional literature. In the empirical part, I chose qualitative research through the interviewing method using the semi-structured interview technique. The research sample consisted of eight clients of the infectious disease ward treated for or shortly after chronic hepatitis C. The data analysis was evaluated by the data categorization method. The results showed a very demanding, mainly physically, course of treatment with a significant impact on the mental and social function of the patient. The reason why the clients had turned to addictive substances was varied, from a curiosity in youth, through addiction as a consequence of trauma from sexual abuse in childhood, to addiction as a consequence of leaving a children's home without the adequate social preparedness and assistance. The components of the multidisciplinary team that intervened the most were healthcare, psychiatry, psychology - psychotherapy, the District Social Security Administration, and the Labour Office. This work identifies the issues that are not widely spoken of. It has shown us that not only addiction to addictive substances is dangerous for the individuals, but that the health problems do not end even after the treatment of their addiction, and the clients have to undergo a rather demanding treatment in order to eliminate these problems. However, on a society-wide level, it is desirable that as many of these clients as possible be treated with a positive result, even if they are drawing large amounts from the general health insurance. The results could serve as materials for the prevention of socio-pathological phenomena, for educational activities in schools or for institutions in, for example, K-centres, which provide help to addictive substance users.
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The diagnostics of the hepatitis C virus with usage of methods of the molecular biology
JAKUBCOVÁ, Markéta
Hepatitis C is a virus caused liver irritation, which leads to hepatocelular carcinoma in more than a quarter of cases. Six different genotypes with at least fifty different subtypes are known (3). Hepatitis C virus rapidly undergoes mutation, which causes every infected person to have more of slightly different variants of the virus in their body. Consequently, the immune system often loses control over the virus, which can lead to the observed chronicity. Qualitative confirmation is important for determination of the presence of the virus. Quantitative confirmation, on the other hand, is crucial for commencing and monitoring of the treatment. (20) The most often examined species is blood serum, or plasma. HCV can be diagnosed directly, or indirectly. Confirmation of the presence of anti-HCV antibodies (indirectly) by Elisa method is routine and very basic screening in most laboratories. Detectable anti-HCV antibodies occur three weeks to six months after the primoinfection (every person?s immune system reacts differently). This indirect test is usually negative in early phases of infection. (21) For direct confirmation of the virus, solely PCR ? polymerase chain reaction ? methods are used. In the first phase, it is necessary to isolate RNA of the virus from the biological material. This can be done either by a special isolation instrument, or manually, using special isolating kits. Use of different methods may result in different yield and purity of RNA. In my work, I used only hand column isolation. After obtaining the RNA of the virus, qualitative confirmation follows. In my case, an ?In house? method was used ?the RT PCR (reverse transcription PCR) followed by the PCR amplification of specific segment of viral RNA, which are visualized by gel electrophoresis in presence of ethidium bromide, which works as an intercalation species. Double step PCR is used. In the first step, reverse transcription progresses for 30 minutes at 50°C. In the presence of RNase inhibitor and reverse transcriptase enzyme RNA is overwritten to cDNA, which is subsequently multiplicated using specific primers. This process is a part of one PCR reaction, which takes place in the Thermocycler in the presence of specific primers, water, buffer, nucleotides, and specific enzyme. For the second round of Nested PCR two different sequences of primers are used. PCR is used for enhancing the sensitivity and specifity of the method. Products of the PCR are then visualized on the agarose gel with intercalation agent (most commonly ethidium bromide).(3) Quantitative determination of viremia level is carried out with Real Time PCR. Detection proceeds in real time using fluorescence dyes. Dyes are usually bonded onto oligonucleotide probe, which specifically bonds to a PCR amplificate. Measuring the intensity of fluorescence enables confirmation and quantification of the products. The system is also able to identify potencial PCR inhibition with internal control. After addition of standards of known concentration, we can work out a calibration curve and use it to quantify measured samples (19). In this work, two isolation kits are compared: a) High Pure Viral RNA Kit (Roche) ? kit used for isolation viral RNA from biological material (serum, plasma). In the first phase, biological material is decomposed, RNA is collected on the column with membrane, washed multiple times and eluted to water for use in Real Time PCR (8) b) QIAamp Viral RNA mini Kit (Qiagen) ? kit used for isolation of viral RNA from blood plasma, or serum. Firstly, biological material is decomposed, RNA is collected on the column with membrane, washed multiple times and eluted to water for use in Real Time PCR (13).The samples isolated with the QIAamp Viral RNA Kit (Qiagen) have shown higher values of measured viral RNA after quantification.
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The Awareness of Hepatitis B, C, D and Possible Ways of its Transmission at Intravenous Drug Users in Pardubice Region
TICHÁ, Žaneta
This thesis deals with the knowledge regarding viral hepatitis B, C and D considering intravenous drug application. Viral hepatitides represent one of big medical problems around the world. Hepatitis B, C and D belong among the most serious infectious diseases blood and sexually transmitted. However, these are not the only possible ways of transmission. At viral hepatitides there exists a risk of infection not always due to their own fault. We are all exposed to this risk and therefore it is important to inform and raise awareness about viral hepatitides not only among intravenous drug users but also general population. All over the world 2 billion of people are estimated to suffer viral hepatitis B, over 180 million hepatitis C and the number of people infected with viral hepatitis D is estimated for 15 million. Viral hepatitides can cause acute illness as well as we must take into account asymptomatic form of this disease. The infected itself does not have to know about his/her disease for a long time, therefore all the stated numbers are estimations. The theoretical part deals with actual situation ? in the first part there is defined the term intravenous drug user and there are listed the most important risks of this way of drug application. Next part is divided into five subchapters. In the first three subchapters I deal with viral hepatitides B, C and D. These subchapters describe an occurrence, source of infection, ways of transmission and symptoms of individual diseases. The second last subchapter describes prevention of viral hepatitides and the last one presents a treatment of viral hepatitides. Practical part includes a research. The research sample created intravenous drug users attending Kontaktní centrum Laxus o. s. (Contact Centre) in Pardubice and a field program in the Pardubice region. For this research I chose the method of questioning, the questionnaire technique. The result data were processed and consequently entered into graphs and tables. The results were assessed and compared to an available literature within a discussion. There were defined two targets and six hypotheses within the research.
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