Made available in DSpace on 2015-04-20T12:31:40Z (GMT). No. of bitstreams: 1 Tese Final Luciana Botinelly.pdf: 2352589 bytes, checksum: 2dfaff58148566cdae4e7f8988cc79f7 (MD5) Previous issue date: 2010-10-27 / Fundação de Amparo à Pesquisa do Estado do Amazonas / In almost all Tuberculosis control programs in the world, sputum smear analyze is the first choice diagnosis tool of the disease. The World Health Organization estimates that 150 million people, between the years 2002 and 2020, would have Tuberculosis, and this would be the minimum number of sputum smears to be performed during this period. The most widely stain used in smear staining is Ziehl-Neelsen s, described for over 125 years. This stain is based on the property that mycobacterias have, including Mycobacterium tuberculosis main causing tuberculosis, to withhold a dye solution composed of fuchsin and phenol (carbolfuchsin) in its cell wall, even after washing with acid-alcohol solution. However, carbolfuchsin is recognized as a carcinogenic substance in rodents, when introduced into diet. There is reference to danger of great harm to human health by prolonged exposure, with irreversible consequences. It is flammable, but heating is recommended for implementation of main coloring technique, which releases combustion gases and vapors hazardous if inhaled. No other less toxic substance was proposed yet. Thus, experimental study of evaluation of twelve alternate dyes on coloring smears was carried out, aimed at implementing in clinical samples suspected of Tuberculosis. In-test dyes solutions were prepared, at different concentrations and without phenol addition. For testing, positive acid-fast bacilli sputum smears and clinical isolate of M. tuberculosis smears were stained without heating, for 5 minutes, bleached with alcohol-acid solution for 1 minute, and counterstained with methylene blue, for 1 minute. The microscopic readings were held in transmitted light and in fluorescence (three filters), at immersion objective (100 x). The most promising dye was the alkaloid from vegetable species (AEV), which allowed fluorescence reading, at green and blue filters, without excessive aggregation to background cells and/or to mucous material present in sputum samples. Sensitivity and specificity tests, and comparison with current bacteriological method confirmed the potential use of AEV. Optimization tests for traditional microscopy reading were made with no success, a fact that indicates other testing. Three purified alkaloids and four alkaloids fractions from amazonian plant species were tested and considered promising for use in mycobacterial stain. They could be added to AEV, providing the ideal dye for Tuberculosis diagnosis, allowing both traditional and fluorescence microscopy. / Em quase todos os programas mundiais de controle da Tuberculose, a avaliação baciloscópica é adotada como primeira opção de exame diagnóstico da doença. A Organização Mundial de Saúde estima que 150 milhões de pessoas, entre os anos de 2002 e 2020, adoecerão por Tuberculose, e este seria o número mínimo de baciloscopias a serem realizadas neste período. A coloração mais utilizada na baciloscopia é a de Ziehl-Neelsen, descrita há mais de 125 anos. Tem como princípio a propriedade que as micobactérias possuem, incluindo o Mycobacterium tuberculosis principal causador da Tuberculose, de reter uma solução corante composta por fucsina básica e fenol (carbolfucsina) em sua parede celular, mesmo após lavagem com solução álcool-ácida. Entretanto, a carbolfucsina é considerada indutora de tumores em roedores, quando introduzida em sua dieta, e há referência a perigo de grande dano para saúde humana em caso de exposição prolongada, com possíveis efeitos irreversíveis. É produto combustível, que em caso de aquecimento, como o recomendado para a execução da principal técnica de coloração, forma gases de combustão e vapores perigosos para inalação. Nenhuma opção de substituto menos tóxico que a carbolfucsina foi até o momento proposto. Desta forma, foi realizado estudo experimental de avaliação de doze corantes alternativos na execução de coloração baciloscópica para micobactérias, visando aplicação em amostras clínicas submetidas ao diagnóstico laboratorial da Tuberculose. Para tanto, foram preparadas soluções dos corantes em teste - em diferentes concentrações e sem adição de fenol. Para os testes, baciloscopias positivas para BAAR, obtidas a partir de amostras de escarro e de isolado clínico de M. tuberculosis, foram coradas sem aquecimento, por 5 minutos, descoradas com solução álcool-ácido por 1 minuto, e contra-coradas com azul de metileno, por 1 minuto. As leituras microscópicas foram realizadas em campo claro e em fluorescência (três filtros), objetiva de imersão (aumento de 100x). O corante mais promissor foi o alcalóide de espécie vegetal (AEV), o qual permitiu visualização bacilar, em microscopia de fluorescência, filtros azul e verde, sem agregação excessiva às células de fundo e/ou à secreção mucóide presente em amostras de escarro. Testes de sensibilidade de detecção bacilar, de especificidade, e comparativo com método bacteriológico tradicional confirmaram o potencial de utilização do AEV. Testes de otimização visando sua utilização para leitura em microscopia tradicional foram realizados, sem obter sucesso, fato que indica a realização de novos testes. Três alcalóides purificados e quatro frações alcaloídicas de possíveis fontes amazônicas do AEV foram testados e considerados como promissores para uso na coloração de micobactérias. Estes podem ao serem agregados ao AEV, propiciar o corante ideal para o diagnóstico da Tuberculose, permitindo visualização tanto à microscopia tradicional, quanto à de fluorescência.

Thesis (MPH.) -- University of Limpopo, 2017. / The present study presents data about the prevalence and determinants of sputum smear non-conversion in smear positive tuberculosis patients. Despite the intervention by the Sekhukhune District Department of Health through continual training and workshops of professional nurses in respect of the NTCP, there are still more challenges observed in terms of TB management. Aim: To investigate the prevalence and determinants of sputum smear non-conversion in smear positive PTB patients after intensive phase of treatment. Method: Quantitative, descriptive retrospective study of TB records was conducted. Data collection was done by extracting data from ETR.net and exporting it to excel. Data cleaning was done before analysis. Data analysis was done using the computer Statistical Package Software for Social research (SPSS) volume 23.1. Findings: 834 TB patients’ records were extracted from the ETR.net database. 34% of records were available at 2 – months; 57% of the patients were males; also, 81% of the patients were diagnosed/treated at PHC facilities; 52% of the patients were HIV positive; 69% percent of the patients who were smear positive grading p+++ failed to convert after two months. In the univariate logistic regression patients with age 20 – 29 were observed to be 4.9 times likely (O.R. = 4.97) to be sputum positive (P = 0.142).Sputum grade 3(p+++) at the time of diagnosis was found to be significantly associated (P = 0.031) with sputum non – conversion after intensive phase of treatment. Conclusion: Two month sputum smear non-conversion is associated with pre-treatment sputum smear grading.

Magister Public Health - MPH (Public Health) / Over the past two decades, despite a growing tuberculosis (TB) epidemic, the South African health system and National TB Programme (NTP) have taken significant steps to ensure improved clinical awareness, early diagnosis, prompt treatment initiation and follow-up of treatment outcomes in cases of TB. The effects of these programmatic measures over time on changes in the severity of disease and presenting clinical profile of patients with pulmonary TB have not been studied. Doing so may provide another window on the impact of TB control initiatives in South Africa.