O presente trabalho apresenta o resultado de uma investigação realizada com 431 doentes de tuberculose pulmonar, internados em 4 Hospitais de Campos do Jordão, São Paulo, Brasil, no ano de 1977, para verificar o conhecimento dos doentes sobre a doença. A coleta dos dados foi feita por meio de entrevista, utilizando-se um formulãrio semi-estruturado contendo 21 perguntas. O trabalho foi realizado com doentes internados pela primeira vez e com reinternados. Na discussão dos dados foi adotada também a distribuição dos doentes em orientados e não orientados. Os resultados mostram que a população em estudo apresenta conhecimentos insuficientes em alguns aspectos importantes da doença, o que justifica as recomendações apresentadas como subsídios para a integração da educação em saúde aos progranas que visam o controle da tuberculose. / This paper reports an investigation carried out in 1977 in Brazil to assess the knowledge that tuberculosis hospital patients have about their disease. Data was collected through interviews based on a total of 21 questions, and grouped into 4 categories: newly hospitalized patients and relapses; patients who received information on the disease and patients who did not. Findings disclosed that patients lacked knowledge about some important aspects of the disease pointing to the need of integration of health education to aimed at the control of tuberculosis.

Submitted by Fabiana Gonçalves Pinto (benf@ndc.uff.br) on 2015-12-11T11:58:50Z No. of bitstreams: 1 Ingrid Ramos Reis Couto.pdf: 671738 bytes, checksum: 69fe2d027ff4b1e498d49a467ae90e32 (MD5) / Made available in DSpace on 2015-12-11T11:58:50Z (GMT). No. of bitstreams: 1 Ingrid Ramos Reis Couto.pdf: 671738 bytes, checksum: 69fe2d027ff4b1e498d49a467ae90e32 (MD5) Previous issue date: 2012 / Mestrado Profissional em Enfermagem Assistencial / Problema: O aparecimento de casos de adoecimentos por tuberculose entre os funcionários do HUAP. Objetivos: Analisar os fatores de risco para tuberculose e o perfil epidemiológico dos funcionários do HUAP/UFF com resultado da prova tuberculínica ≥ 10 mm no período de 2007 a junho de 2011; - Investigar os casos de adoecimento por tuberculose em funcionários do HUAP/UFF no período de janeiro de 2004 a julho de 2011; - Identificar a possível associação entre o perfil epidemiológico dos funcionários que apresentaram positividade na prova tuberculínica e os fatores de riscos para tuberculose. Métodos: trata-se de um survey interseccional, tendo como análise estatística a razão de chances (OR) e análise multivariada dos profissionais que apresentaram conversão na prova tuberculínica. Resultados: foram identificados 10 casos notificados de adoecimentos por tuberculose nos funcionários do HUAP/UFF no período de janeiro de 2004 a julho de 2011, sendo a predominância no sexo masculino com 80%, na qual o perfil de adoecimentos foi de funcionários que não realizavam suas atividades diretamente ao paciente com TB. Os funcionários que apresentaram positividade na prova tuberculínica no período junho de 2007 a março de 2011 teve como variável idade ≥ 50 anos maior risco de conversão na PT com p- valor (0,003), funcionários com tempo de serviço ≥ 30 anos tiveram (OR) 92,3% mais chances de conversão . Funcionários que tinha como ocupação atividades exercidas na categoria laboratório apresentou um risco de conversão 2,2 ( OR) vezes maior quando comparado as demais categorias. Conclusão: foi possível neste estudo estabelecer a relação entre a atividade profissional e a exposição ao risco de contágio em tuberculose. Portanto cabe as chefias de cada setor que as atividades educativas sejam realizadas de forma descentralizada com o objetivo de focar a individualidade de cada setor / Problem: the appearance of cases of illnesses due to tuberculosis among workers of the HUAP. Objectives: analyze risk factors for tuberculosis and epidemiological profile of workers HUAP / UFF results with the tuberculin skin test ≥ 10 mm in the period 2007 to June 2011 - Investigate cases for tuberculosis in workers of HUAP / UFF in From January 2004 to July 2011 - Identify the possible association between the epidemiological profile of workers who were positive in the tuberculin test and the risk factors for tuberculosis. Methods: this is an intersectional survey, with the statistical analysis the odds ratio (OR) and multivariate analysis showed that conversion of the workers in the tuberculin test. Results: were identificad 10 reported case of TB illnesses in workers of HUAP / UFF from january 2004 to july 2011, the predominance in males, with 80%, in which the profile of workers sickness was not performed activities directly to the patient with TB. Workers who tested positive in the tuberculin test in the period june 2007 to march 2011 was variable age ≥ 50 years old increased risk of conversion in PT with p-value (0.003), length of service workers with ≥ 30 years were 92.3% ( OR) conversion as likely . Workers who had occupation activities performed in the category laboratory presented a risk of conversion 2.2 (OR) times higher compared to other categories. Conclusion: this study it was possible to establish the relationship between professional activity and exposure to the risk of contagion in tuberculosis. Therefore it is the heads of each sector that educational activities are conducted in a decentralized manner in order to focus on the individuality of each sector

Bakgrund: Vård av patienter med smittsam lungtuberkulos har blivit allt vanligare. Ett antal fall där personal smittats har förekommit i landet under de senaste åren. Många av dessa patienter har en annan etnisk bakgrund och talar ett annat språk än personalen. Syfte: Att studera sjuksköterskors upplevelser av att vårda patienter med lungtuberkulos på infektionsklinik. Metod: Semi-strukturerade intervjuer genomfördes med sex sjuksköterskor på en infektionsklinik. Materialet analyserades med kvalitativ innehållsanalys. Resultat: Tre kategorier skapades; Sjuksköterskans upplevelse av vårdandet, Sjuksköterskans upplevelse av kommunikationen, Sjuksköterskans upplevelser och tankar kring smittspridning. Sjuksköterskorna upplevde att skillnader utifrån språk och etnisk bakgrund gjorde det svårt att ge den vården de önskade, vilket ledde till frustration och stress. Sjuksköterskorna upplevde glädje och tillfredställelse när de upplevde att de kunde ge god vård och att patienten var nöjd. Avvikelser från riktlinjer och rutiner gav upphov till en oro för smittspridning. Resultatet diskuteras utifrån Hendersons omvårdnadsdefinition. Slutsats: Det behövs bättre kunskaper om interkulturell omvårdnad, och ökade möjligheter till användning av tolk för att förbättra vården av patienterna. Tydliga riktlinjer avseende handläggning av patienterna med lungtuberkulos som är kända och efterföljs av all personal behövs för att öka tryggheten. Att kunna ge en god vård ger sjuksköterskorna en känsla av tillfredställelse i arbetet. / Background: Caring for patients with infectious pulmonary tuberculosis has become increasingly common. A number of cases of employees being infected have occurred in the country in recent years. Many of the patients have a different ethnic background and speak a language other than the staff. Aim: Study nurses experiences of caring for patients with pulmonary tuberculosis at a clinic for infectious diseases. Method: Semi-structured interviews were conducted with six nurses at a clinic for infectious diseases. The material was analysed with qualitative content analysis. Results: Three categories was created; Nurse's experience of caring, nurse's experience of communication, the nurse experiences and thoughts about the spread of infection. The nurses felt that differences based on language and ethnic background made it difficult to provide the care they wanted, leading to frustration and stress. The nurses experienced joy and satisfaction when they felt that they could provide good care and the patient was satisfied. Deviations from the guidelines and procedures gave rise to fears of contamination. The results are discussed on the basis of Henderson's nursing definition. Conclusion: We need a better understanding of cross-cultural care, and increased opportunities for the use of interpreters to improve patient care. Clear guidelines for management of patients with pulmonary TB who are known and followed by all staff needed to increase security. Being able to provide good care gives nurses a feeling of satisfaction at work.

Introdução: O teste Xpert MTB / RIF está sendo cada vez mais utilizado em muitos países como diagnóstico inicial para a tuberculose (TB). Poucos estudos avaliaram o impacto do Xpert no diagnóstico em rotinas de programas de controle de TB no Brasil. O objetivo do presente estudo foi avaliar o impacto da introdução do Xpert MTB / RIF no diagnóstico de TB em uma cidade com alta incidência de TB no Brasil. Métodos: Incluímos pacientes avaliados com testes diagnósticos convencionais durante um ano antes da introdução do Xpert (grupo pré-Xpert) e pacientes avaliados usando Xpert durante um ano após a introdução do teste (grupo pós-Xpert). Resultados: 620 pacientes preencheram os critérios de inclusão (208 no grupo pré-Xpert e 412 no grupo pós-Xpert) e foram incluídos na análise. O tempo até o diagnóstico de TB foi menor no grupo pós-Xpert (0,7 dias, IQR: 0,5-1,0 dias) do que no grupo pré-Xpert (2,0 dias, IQR: 2,0-2,0 dias) (p <0,0001). Características atípicas da doença, como menor perda de peso, febre, dispneia, sudorese noturna e hemoptise; baciloscopia de escarro negativa; cultura negativa e radiografia de tórax atípica de TB foram mais comuns no grupo pós-Xpert do que no grupo pré-Xpert (p <0,0001 para todos). Conclusões: Observamos que a implementação do ensaio Xpert MTB / RIF, em rotinas de programas de controle de TB, melhora e facilita o diagnóstico de tuberculose, especialmente nos casos com manifestações da doença atípica. Esses resultados podem provavelmente ser generalizados para locais com incidência de TB similar. / Introduction: The receptor for advanced glycation end products (RAGE) is expressed in normal lungs and is upregulated during inflammation and infection. The interaction between AGEs and RAGE on the plasma membrane causes oxidative stress and apoptosis in lung cells. The objective of this study is to evaluate plasma levels of AGEs and its soluble receptor (sRAGE) in patients with active TB and healthy controls, and to investigate their relationship with food intake and nutritional status. Methods: Case-control study. AGE (carboxymethil lysine, CML) and RAGE were measured by Elisa. Nutritional assessment was performed by body mass index, triceps skin-fold thickness, mid-arm circumference, mid-arm muscle circumference, bioelectrical impedance analysis, and food frequency questionnaire. Results: 35 TB patients and 35 controls were included in the study. The mean S-RAGE levels were higher in TB patients than in controls (68.5 ± 28.1 vs 57.5 ± 24.0, p=0.046). Among cases that were current smokers, lower S-RAGE levels were associated with mortality (S-RAGE levels= 58.0 ± 36.5 [non-survivors] vs 71.3 ± 25.6 [survivors], p=0.006), and with weight loss (S-RAGE levels= 65.6 ± 27.4 [weight loss] vs 98.6 ± 16.7 [no weight loss], p=0.034). There was no statistically significant difference in CML levels and diet CML content between cases and controls. Malnutrition was more frequent in cases than in controls, but there was no correlation between nutritional parameters and CML or S-RAGE levels. Conclusions: TB patients had higher S-RAGE levels than controls. S-RAGE may play a role in disease manifestations and outcomes, being associated with weight loss and mortality.

Thesis (PhD)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: The immune response against M. tuberculosis is multifactorial, involving a network of innate and adaptive immune responses. Characterization of the immune response, a clear understanding of the dynamics and interplay of different arms of the immune response and the identification of infection-stage specific biomarkers are critical to allow the development of better tools for combating tuberculosis. In an attempt to identify such biomarkers, we studied pulmonary tuberculosis patients and their contacts in Addis Ababa, Ethiopia as part of EDCTP and BMGF funded tuberculosis projects by using multiplex techniques. We analysed 45 genes using the Multiplex Ligation Dependent Probe Amplification (MLPA) technique and the expression of IL-4δ2, BLR1, MARCO, CCL-19, IL7R, Bcl2, FcyR1A, MMP9, and LTF genes discriminate TB cases from their healthy contacts. FoxP3, TGFß1 and CCL-19 discriminate latently infected from uninfected contacts. Single genes predict with an area under the Receiver Operating Characteristic (ROC) curve of 0.68 to 0.85 while a combination of genes identified up to 95% of the different groups. Similarly, the multiplex analysis of cytokines and chemokines also showed that single or combinations of plasma cytokines and chemokines discriminate between different clinical groups accurately. The median plasma level of EGF, fractalkine, IFN-y, IL-4, MCP-3 and IP-10 is significantly different (p<0.05) in active tuberculosis and non active tuberculosis infection and the median plasma levels of IFN-y, IL-4, MCP-3, MIP-1ß and IP-10 were significantly different (p<0.05) before and after treatment. We also found a significant difference (p<0.05) in plasma levels of cytokines of patients infected with the different lineages and different families of the modern lineage. The plasma level of IL-4 was significantly higher in patients infected with lineage 3 (p<0.05) as compared to lineage 4 and the CAS familyinfected patients had a higher plasma level of IL-4 (P<0.05) as compared to patients infected with H and T families but there was no difference between H and T families. We identified genes and cytokines which had been reported from other studies in different settings and we believe that these molecules are very promising biomarkers for classifying active tuberculosis, latent infection, absence of infection and treated infection. These markers may be suitable for the development of clinically useful tools but require further validation and qualification in different populations and in larger studies. / AFRIKAANSE OPSOMMING: Die immuunrespons teen M. tuberculosis is multifaktoriaal en betrek ‘n netwerk van niespesifieke and spesifieke immuunresponse. Karakterisering van die immuunrespons, ‘n duidelike insig in die dinamika en tussenspel deur die verskillende arms van die immuunrespons en die identifikasie van spesifieke biomerkers is krities belangrik om die ontwikkeling van nuwe hulpmiddels teen tuberkulose te bevorder. In ‘n poging om sulke biomerkers te identifiseer het ons pulmonale tuberkulose pasiënte en hulle kontakte in Addis Ababa, Etiopië, as deel van die EDCTP en BMGF befondste tuberkulose projekte bestudeer met multipleks tegnieke. Ons het 45 gene analiseer met ‘Multiplex Ligation Dependent Probe Amplification (MLPA)’ en gevind dat die geenuitdrukking van IL-4•2, BLR1, MARCO, CCL-19, IL7R, Bcl2, Fc•R1A, MMP9, en LTF TB pasiënte van hulle kontakte onderskei. FoxP3, TGF•1 en CCL-19 onderskei tussen latent infekteerde en ongeïnfekteerde kontakte. Enkele gene voorspel met ‘n area onder die ‘Receiver Operating Characteristic (ROC)’ kurwe van 0.68 tot 0.85 terwyl die kombinasie van gene 95% van die verskillende groepe identifiseer. Soortgelyk het multipleks analise van sitokiene en chemokiene verskillende kliniese groepe akkuraat van mekaar onderskei. Die mediane plasmavlakke van EGF, fractalkine, IFN-•, IL-4, MCP-3 en IP-10 is beduidend verskillend (p<0.05) in aktiewe tuberkulose en nie-aktiewe tuberkulose infeksie en die mediane plasmavlak van IFN-•, IL-4, MCP-3, MIP-1• en IP-10 was beduidend verskillend voor en na behandeling. Ons het ook beduidende verskille (p<0.05) in plasmavlakke van sitokiene in pasiënte gevind wat infekteer is met verskillende stamme and verskillende families van die moderne stamme. Die plasmavlak van IL-4 was beduidend hoër in pasiënte wat infekteer is met stam 3 (p<0.05) teenoor stam 4 en die CAS familie-infekteerde pasiënte het ‘n hoër plasmavlak van IL-4 (p<0.05) teenoor pasiënte met H en T familie infeksie hoewel daar geen versikke was tussen die H en T families nie. Ons het gene en sitokiene identifiseer wat deur ander werkers onder verskillende omstandighede ook beskryf is en ons glo dat hierdie molekules baie belowende biomerkers is om aktiewe tuberkulose, latent tuberkulose, die afwesigheid van infeksie en behandelde infeksie van mekaar te onderskei. Hierdie merkers mag toepaslik wees vir die ontwikkeling van bruikbare kliniese hulpmiddele maar benodig verdere validasie en kwalifikasie in verskillende populasiegroepe en in groter studies. / Bill and Melinda Gates Foundation (BMGF) / European and Developing Countries Clinical Trials Partnership (EDCTP) / African European Tuberculosis Consortium (AE TBC).

Thesis (MScPhysio)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: Background: Pulmonary tuberculosis (PTB) remains a major concern worldwide. Although PTB is curable, both the disease and its treatment may have considerable medical, social and psychological consequences which may result in a decreased quality of life and functioning. Characterization of the functional capabilities of PTB patients post-treatment and the impact of PTB on their quality of life may identify a need for more holistic management of PTB treatment that extends beyond microbiological cure. Methods: Firstly, an in-depth scoping review was conducted using the following key words: Pulmonary tuberculosis (MESH term) and Health related quality of life (HRQoL), Pulmonary tuberculosis (MESH term) and Spirometry and Pulmonary tuberculosis (MESH term) and Six minute walk test or 6MWT to review the current literature reporting on the HRQoL, lung function measurements and exercise capacity of a PTB population (Chapter 2). Secondly, a cross-sectional, quantitative, descriptive study was conducted. The study setting included five primary health care facilities (PHCF) in the Breede Valley sub-district of the Cape Winelands East District, Western Cape, South Africa. Adult patients diagnosed with PTB, 18 years and older and who were successfully managed through the Cape Winelands District Health Care system were considered for the study if they had least two negative sputum sample results and had completed at least five months of anti-tuberculosis treatment. Post treatment bronchodilator lung function tests, health related quality of life using the BOLD core questionnaire and six minute walk test distance (6MWD) was measured. Findings: The comprehensive broad search of the literature yielded a total of 2446 articles. A total of 2422 articles were excluded since the title; abstract or full text article did not conform to the review question or were eliminated as duplicates across databases. Twenty-seven articles divided amongst the three subsections i.e. PTB and HRQoL (n=13), PTB and Spirometry (n=9) and PTB and exercise capacity (n=6), were included in the review. In the cross-sectional study, 328 names were obtained from the TB registers of the five included PHCF of which 45 patients were included in the study (56% male; mean age, 39.88±10.20 years). The majority of patients (n= 206; 63%) were not contactable, and could not be recruited. Approximately half the total sample, (n=23; 52%) presented with normal lung function while n=11 (25%) presented with a restrictive pattern, n=9 (21%) presented with an obstructive pattern and only n=1 (2%) presented with a mixed pattern (defined as FEV1<80% predicted, FVC<80% predicted and FEV1/FVC<0.7). The mean six minute walk distance (6MWD) was 294.5m±122.7m. Respondents scored poorly on all sub-domains of the SF-12v2 except vitality. Role emotional and role physical scored lowest with mean scores of 28.1 and 35.27 respectively, while vitality scored the highest with 52.78. Stellenbosch University https://scholar.sun.ac.za 4 | P a g e Conclusion The findings of this thesis suggest that even after microbiological cure, PTB patients may suffer from a decreased quality of life, impaired lung function and a decreased exercise capacity. Specific challenges to data collection in a rural region were identified; which included patient recruitment, field testing of exercise capacity (6MWD), and the generalizabilty of standardized questionnaires in rural regions. The findings of this pilot study serves to inform the planning of a larger observational study, in the rural Cape Winelands of the Western Cape, South Africa. / AFRIKAANSE OPSOMMING: Agtergrond Pulmonêre tuberkulose (PTB) wek wêreldwyd steeds groot kommer. Hoewel dit geneeslik is, kan die siekte sowel as die behandeling daarvan beduidende mediese, maatskaplike en sielkundige gevolge hê, wat lewensgehalte en funksionering kan knou. Die tipering van PTB-pasiënte se funksionele vermoëns ná behandeling sowel as die impak van PTB op hul lewensgehalte kan dalk dui op ’n behoefte aan die meer holistiese bestuur van PTB-behandeling, wat méér as blote mikrobiologiese genesing insluit. Metodes Eerstens is ’n diepgaande bestekstudie aan die hand van die volgende trefwoorde onderneem: pulmonêre tuberkulose (MeSH-term) en gesondheidsverwante lewensgehalte (HRQoL), pulmonêre tuberkulose (MeSH-term) en spirometrie, en pulmonêre tuberkulose (MeSH-term) en die ses minute lange stapafstandtoets (6MWT). Na aanleiding daarvan is die huidige literatuur oor die HRQoL, longfunksiemetings en oefenvermoë van ’n PTB-populasie bestudeer (hoofstuk 2). Tweedens is ’n kwantitatiewe, beskrywende deursneestudie onderneem. Die studie-omgewing het bestaan uit vyf fasiliteite vir primêre gesondheidsorg in die Breedevallei-subdistrik van die streek Kaapse Wynland-Oos, Wes-Kaap, Suid-Afrika. Volwasse pasiënte van 18 jaar en ouer wat met PTB gediagnoseer is en suksesvol deur die distriksgesondheidsorgstelsel van die Kaapse Wynland-streek bestuur word, is vir die studie oorweeg indien minstens twee van die pasiënt se sputummonsters TB-negatiewe resultate opgelewer het en die persoon reeds minstens vyf maande vir tuberkulose behandel is. Studiemetings het ingesluit brongodilator-longfunksietoetse ná behandeling, gesondheidsverwante beoordelings van lewensgehalte met behulp van die BOLD-vraelys, en die aflegging van ’n ses minute lange stapafstandtoets (6MWT). Bevindinge Die omvattende breë soektog van die literatuur het 'n totaal van 2446 artikels opgelewer. 'n Totaal van 2422 artikels is uitgesluit, aangesien die titel; abstrakte of volledige teks artikel het nie voldoen aan die navorsings vraag, of is uitgeskakel as duplikate oor databasisse. Sewe en twintig artikels verdeel tussen die drie onderafdelings, naamlik PTB en HRQoL (n = 13), PTB en Spirometrie (n = 9) en PTB en oefening kapasiteit (n = 6), is ingesluit in die oorsig. In die deursneestudie is 328 name uit die TB-registers van die vyf ondersoekpersele bekom. Altesaam 45 pasiënte (56% mans; gemiddelde ouderdom 39.88±10.20 jaar) is by die studie ingesluit. Die oorgrote meerderheid pasiënte (n = 206; 63%) kon nie bereik word nie, en dus ook nie gewerf word nie. Ongeveer die helfte van die algehele steekproef (n = 23; 52%) se longfunksie was normaal; n = 11 (25%) het ’n restriktiewe patroon getoon; n = 9 (21%) ’n obstruktiewe patroon, en slegs n = 1 (2%) ’n gemengde patroon (wat omskryf word as ’n FEV1-voorspellingswaarde van <80%, ’n FVC-voorspellingswaarde van <80%, en FEV1/FVC van <0.7). Die gemiddelde afstand wat in die ses minute lange staptoets afgelê is (6MWD), was 294,5 m±122,7 m. Respondente behaal swak Stellenbosch University https://scholar.sun.ac.za 6 | P a g e op al die sub-domein van die SF-12v2 behalwe vitaliteit. Rol emosionele en rol fisiese behaal laagste met die gemiddelde tellings van 28.1 en 35,27 onderskeidelik, terwyl vitaliteit behaal die hoogste met 52,78. Gevolgtrekking Die bevindinge van hierdie tesis gee te kenne dat PTB-pasiënte selfs ná mikrobiologiese genesing dalk swakker lewensgehalte, verswakte longfunksie en ’n afname in oefenvermoë ondervind. Bepaalde uitdagings vir data-insameling in ’n landelike omgewing is uitgewys, onder meer pasiëntewerwing, veldtoetsing van oefenvermoë (6MWD) en die veralgemeenbaarheid van gestandaardiseerde vraelyste in landelike gebiede. Die bevindinge van hierdie proefstudie kan gebruik word om die beplanning van ’n groter waarnemingstudie in die landelike Kaapse Wynland-streek in die Wes-Kaap, Suid-Afrika, te rig.

Thesis (MScMedSc)--Stellenbosch University, 2012. / Bibliography / ENGLISH ABSTRACT: South Africa is ranked amongst the top tuberculosis (TB) burden countries in the world and the Western Cape has a particularly high incidence of the disease. Previous studies have showed that several genes may play crucial roles in susceptibility to TB. In this study, we investigated the role of three genes previously associated with susceptibility to TB and progression to disease. These genes were Surfactant protein D (SFTPD), Macrophage receptor with collagenous structure (MARCO) and CD14. The proteins from these genes bind M. tuberculosis and are involved in the uptake of the bacteria into macrophages. The study investigated the role of ten polymorphisms from SFTPD and MARCO within a South African Coloured (SAC) population, where tuberculosis is highly prevalent. A casecontrol study design was used and polymorphisms were genotyped with Taqman® genotyping assays and amplification refractory mutation system polymerase chain reaction (ARMS-PCR). The results were analysed for association with disease, linkage disequilibrium, haplotypes and gene-gene interactions. Allele and genotype frequencies were also determined which allowed for comparisons to other populations. Five SNPs were associated with TB: two in SFTPD (rs1923537; rs2255326) and three in MARCO (rs1318645; rs3943679; rs2119112). The associated SNPs were located in regions other than exons and the effects of polymorphisms in these regions are not well understood but studies in other genes have shown them to play a functional role. Gene-gene interaction analysis showed that polymorphisms interacted with each other within and between genes, illustrating the importance of epistasis and the complexity of the genetic influences on TB. In addition to the case-control association studies, the role of the rs2569190 promoter SNP in CD14 was assessed. Gene-expression analysis was conducted with qPCR and a reporter gene assay and results from both of these approaches showed that individuals with the TT genotype had a twofold greater expression level than individuals with the CC genotype. Previously, the TT genotype has been associated with stronger promoter activity and expression of soluble CD14 in serum. Since the TT genotype was present at a higher frequency in the control group, we speculate that greater expression of CD14 may contribute to a more TB resistant phenotype. The work presented in this study illustrates the importance of the host genetic component of TB. Genetic studies will eventually revolutionize the current treatment regime as the identification of vulnerable individuals and populations will aid in the development of personalised medicines. / AFRIKAANSE OPSOMMING: Suid-Afrika is een van die top tuberkulose (TB) lande in die wêreld en die Wes-Kaap het veral ‘n hoë insidensie van die siekte. Vorige studies het gewys dat verskeie gene bydra to die vatbaarheid vir tuberkulose. In hierdie studie het ons drie gene, wat voorheen vir vatbaarheid vir tuberkulose en progressie na die siekte ondersoek is, bestudeer. Hierdie gene is Surfactant protein D (SFTPD), Macrophage receptor with collagenous structure (MARCO) en CD14. Die proteïene van hierdie gene bind M. tuberculosis en is betrokke in die opname van die bakterieë in die makrofages. Hierdie studie het tien polimorfismes van SFTPD en MARCO in die Suid-Afrikaanse Kleurlingbevolking (SAK), wat ‘n hoë TB insidensie het, getoets. Pasiënt-kontrole assosiasie studies is gedoen en polimorfismes is gegenotipeer met Taqman® genotiperingsisteem en die amplifikasie refraktoriese mutasie sisteem polimerase ketting reaksie (ARMS-PCR). Die resultate is geanaliseer vir assosiasies met TB, koppelings disekwilibrium, haplotipes en geen-geen interaksies. Alleel en genotype frekwensies is ook bepaal en vergelyk met die van ander bevolkings. Vyf enkel nukleotied polimorfismes (ENPs) is met TB geassosieer: twee in SFTPD (rs1923537; rs2255326) en drie in MARCO (rs1318645; rs3943679; rs2119112). Die geassosieerde ENPs was nie in eksons nie. Die effek van polimorfismes in areas anders as eksons word nie goed verstaan nie, maar studies het bewys dat hulle wel ‘n funksionele rol kan hê. Geen-geen interaksie analise het gewys dat polimorfismes interaksies met mekaar binne sowel as tussen gene gehad het, wat die belangrikheid van epistase en die kompleksiteit van genetiese invloede op TB illustreer. Tesame met die pasiënt-kontrole assosiasie studies is die rol van die rs2569190 promoter ENP in CD14 ook ondersoek. Geenuitdrukkingsanalise is gedoen met qPKR en rapporteerder geen toetse. Die resultate van beide hierdie benaderings het gewys dat individue met die TT genotipe twee keer soveel uitdrukkingsvlakke gehad het as individue met die CC genotipe. Die TT genotipe is voorheen geassosieer met sterk promoter aktiwiteit en die uitdrukking van oplosbare CD14 in serum. Aangesien die TT genotipe meer in die kontrolegroep gevind is, spekuleer ons dat die hoër uitdrukking van CD14 kan bydra tot ‘n meer TB weerstandbiedende fenotipe. Hierdie werk illustreer die belangrikheid van die gasheer genetiese komponent in TB. Genetiese studies sal in die toekoms die huidige behandeling regime revolusioneer, aangesien die identifikasie van individue en bevolkings met ‘n hoë risiko om TB te ontwikkel sal bydra tot die ontwikkeling van persoonlike medisynes. / The National Research Foundation (NRF); the South African Medical Research Council (MRC); Stellenbosch University and the Harry Crossley Foundation.

Comparou-se a eficácia da baciloscopia in natura e pós concentração (Coloração Ziehl Neelsen) e cultura tradicional e automatizada MA de amostras de escarro ou saliva/muco para a detecção do Complexo M. tuberculosis (MTB) em Centro de Referencia Ambulatorial para Tuberculose na Cidade de São Paulo. A identificação do grupo MTB baseou-se no crescimento em meio com Ácido para nitrobenzóico, e visualização do fator corda. Do total de 374 amostras, 228 eram de pacientes sob diagnóstico inicial e 146 em tratamento. 83 amostras eram saliva/muco. Destas, sete foram positivas à baciloscopia, 03 (3,6%) no material concentrado e uma (1,2%) in natura (p=0.5 McNemar Test). Cinco amostras (6%) positivas na cultura pelo método tradicional e 07 (8,4%) p=0.5 pelo MA. As amostras de escarro eram 74 foram positivas na baciloscopia, sendo 34 (11,7%) in natura e 45 (15,5%) no material concentrado, p=0.001. No método tradicional 56 (19,2%) e 67 (23%) pelo MA p=0.001. Independente da característica da amostra a baciloscopia pós concentração e a cultura pelo MA foram mais sensíveis. / Effectiveness of bacilloscopy were compared in natura and pos-concentrated (Ziehl Neelsen staining), and tradicional and automated culture-AM of sputum or saliva/mucus samples for detection of M. tuberculosis complex (MTB) in Ambulatory Reference Center in the city of São Paulo. Identification of MTB group was based on growth in medium with para nitrobenzoic acid, and cord factor visualization. From the total 374 samples, 228 were from patients under initial diagnostic and 146 in treatment. 83 samples were saliva/mucus. Seven of these were bacilloscopy positive, 03 (3.6%) in concentrated material and one (1.2%) in natura (p=.500ª McNemar Test). Five samples (6%) positive for culture by traditional method and 07 (8.4%) p=0.5 for AM. For sputum 74 were bacilloscopy positive, being 34 (11.7%) in in natura and 45 in concentrated material, p=0.001. In traditional method, 56 (19.2%) and 67 (23%) for AM, p=0.001. Independently of sample characteristics, pos-concentrated bacilloscopy and culture by AM were more effectiveness.

Comparou-se a eficácia da baciloscopia in natura e pós concentração (Coloração Ziehl Neelsen) e cultura tradicional e automatizada MA de amostras de escarro ou saliva/muco para a detecção do Complexo M. tuberculosis (MTB) em Centro de Referencia Ambulatorial para Tuberculose na Cidade de São Paulo. A identificação do grupo MTB baseou-se no crescimento em meio com Ácido para nitrobenzóico, e visualização do fator corda. Do total de 374 amostras, 228 eram de pacientes sob diagnóstico inicial e 146 em tratamento. 83 amostras eram saliva/muco. Destas, sete foram positivas à baciloscopia, 03 (3,6%) no material concentrado e uma (1,2%) in natura (p=0.5 McNemar Test). Cinco amostras (6%) positivas na cultura pelo método tradicional e 07 (8,4%) p=0.5 pelo MA. As amostras de escarro eram 74 foram positivas na baciloscopia, sendo 34 (11,7%) in natura e 45 (15,5%) no material concentrado, p=0.001. No método tradicional 56 (19,2%) e 67 (23%) pelo MA p=0.001. Independente da característica da amostra a baciloscopia pós concentração e a cultura pelo MA foram mais sensíveis. / Effectiveness of bacilloscopy were compared in natura and pos-concentrated (Ziehl Neelsen staining), and tradicional and automated culture-AM of sputum or saliva/mucus samples for detection of M. tuberculosis complex (MTB) in Ambulatory Reference Center in the city of São Paulo. Identification of MTB group was based on growth in medium with para nitrobenzoic acid, and cord factor visualization. From the total 374 samples, 228 were from patients under initial diagnostic and 146 in treatment. 83 samples were saliva/mucus. Seven of these were bacilloscopy positive, 03 (3.6%) in concentrated material and one (1.2%) in natura (p=.500ª McNemar Test). Five samples (6%) positive for culture by traditional method and 07 (8.4%) p=0.5 for AM. For sputum 74 were bacilloscopy positive, being 34 (11.7%) in in natura and 45 in concentrated material, p=0.001. In traditional method, 56 (19.2%) and 67 (23%) for AM, p=0.001. Independently of sample characteristics, pos-concentrated bacilloscopy and culture by AM were more effectiveness.

Les méthodes pour le diagnostic des infections à mycobactéries sont restées pratiquement inchangées pendant des décennies et n'auraient probablement pas progressé sans la réémergence inattendue de la tuberculose (TB) au cours des vingt dernières années du 20ème siècle. Le diagnostic de laboratoire de la tuberculose pulmonaire repose principalement sur la détection des organismes du complexe Mycobacterium tuberculosis (MTC) dans les prélèvements respiratoires par leur isolement et identification ou par les méthodes moléculaires. Pour les patients qui n’expectorent pas, des échantillons du tractus respiratoire peuvent être obtenus par des procédures invasives. En nous basant sur la survie connue de MTC dans le liquide gastrique, nous avons considéré que les MTC devraient également être détectables dans les selles. Nous avons recherché des MTC en parallèle dans des échantillons de tractus respiratoire et dans les selles récoltés des mêmes patients. Mycobacterium tuberculosis a été détecté après décontamination à la chlorhexidine dans des cultures sur des milieux à base d'œuf et par l'examen direct microscopique après coloration de Ziehl-Neelsen. Après la mise au point d’une technique originale d’extraction de l’ADN total à partir des selles, nous avons utilisé la détection de M. tuberculosis par PCR en temps réel en duplex utilisant le gène IS6110 comme cible moléculaire et un contrôle interne de présence d’inhibiteurs de la PCR. Ces travaux mettent en évidence l’intérêt et la sensibilité de l’analyse des selles pour le diagnostic de la tuberculose pulmonaire. Cette technique est aujourd’hui utilisée dans notre laboratoire en routine et est en cours d’évaluation par d’autres équipes.Actuellement, l'identification des mycobactéries basée sur l’analyse des caractéristiques biochimiques (test à l’uréase, test de la nitrate réductase, test de la résistance à la pyrazinamide (pza), test de la sensibilité, test de la résistance à la thiophen-2-carboxylic acid hydrazide (TCH) etc …) est le plus souvent remplacée par des méthodes de biologie moléculaire dont l’amplification par PCR d'un gène spécifique. Ces méthodes sont laborieuses et peuvent exiger un degré d'expertise élevé. Afin de simplifier l'identification précise des isolats de mycobactéries en laboratoire de routine, après avoir vérifié l’inactivation des MTC par la chaleur et l’éthanol pour pouvoir travailler hors du laboratoire P3, nous avons mis au point les conditions pour l'utilisation de la spectrométrie de masse « matrix-assisted laser desorption and ionisation-time-of-flight» (MALDI-TOF-MS) en association avec un logiciel bioinformatique spécifique, comme outil d’identification et de différenciation entre les membres du complexe MTC, du complexe Mycobacterium avium et les autres mycobactéries non-tuberculeuses. Nous avons ensuite appliqué cette technique afin d’identifier les mycobactéries à partir du milieu liquide Middlebrook 7H10 utilisé dans une étuve automatisée. L’identification des mycobactéries isolées dans notre laboratoire, est maintenant réalisée en routine par MALDI-TOF-MS. L’ensemble de ces travaux contribue à améliorer le diagnostic de routine de la tuberculose pulmonaire et les techniques mises au point sont utilisées en routine dans notre laboratoire, en particulier dans le cadre d’un « kit tuberculose ». / The diagnosis of mycobacterial infections remained practically unchanged during numerous decades and would not probably have progressed of the whole without the unexpected re-emergence of the tuberculosis (TB) during the last twenty years of the 20th century.The diagnosis of laboratory of pulmonary tuberculosis is mainly based on the detection of microorganisms of the Mycobacterium tuberculosis complex (MTC) in the respiratory specimens. For the patients who do not expectorate, samples of the respiratory tract can be only obtained according to invasive procedures. Based on the survival known for MTC organisms in the gastric liquid, we considered that MTC organisms would be detectable also in samples of stools. We compared the presence of bodies MTC in samples of respiratory tract and the specimens of stools collected from the same patients. MTC was detected in cultures on egg-based media after appropriate decontamination using the chlorhexidine and by the microscopic examination after Ziehl-Neelsen staining. After the set on of an original protocol of the total DNA extraction from stools, we were able to use by the detection with the real-time PCR of IS6110 using internal controls as PCR inhibitor control. This protocol illustrate the utility of stool analysis for the diagnosis of pulmonary tuberculosis, this technique is actually used in routine in our laboratory, it is also under investigation by other research teams.Phenotypic identification of mycobacteria based on the analysis of biochemical pattern (urase test, nitrate reductase test, resistance to pyrazinamide (pza) test, susceptibility to thiophen-2-carboxylic acid hydrazide (TCH) test, etc …) is more often replaced by molecular methods using DNA, including the development by PCR of a specific gene. These methods are generally laborious and can require a considerable degree of expertise. To simplify the identification specifies isolates of mycobacteria routinely in the laboratory, we estimated the use of the mass spectrometry "matrix-assisted laser desorption and ionization-time-of-flight" (MALDI-TOF MS) in association with a specific bioIT software, was capable of identifying and of distinguishing between the members of Mycobacterium tuberculosis complex, Mycobacterium avium complex and the non- tuberculosis mycobacteria. We verified the MTC inactivation by heating or by ethanol allowing the analysis of the inactivated MTC out of the P3 laboratory; we studied the cost-effectiveness of this method from the blood solid media by extrapolating this approach in the emergent countries. We then applied the same method to identify mycobacteria from Middlebrook 7H10 liquid media used in an automated oven. Identification of cultured Mycobacteria is now done in routine in our laboratory by MALDI-TOF-MS. These data contribute in improving the routine diagnosis of pulmonary tuberculosis and this protocol is then used routinely in our laboratory, particularly with the “kit de tuberculose” we set on.