Detection of Mycobacterium leprae infection in wild nine-banded armadillos (Dasypus novemcinctus) using the rapid ML Flow test

Mycobaterium leprae infection was investigated in armadillos from the State of Espírito Santo, Brazil. The ML Flow test was performed on 37 nine-banded armadillos and positive results were found in 11 (29.7%). The ML Flow test may be used to identify possible sources of Mycobaterium leprae among wild armadillos.

Nosocomial Mycobacterium tuberculosis transmission among healthcare students in a high incidence region, in Vitória, State of Espírito Santo

This open comparative study for evaluating hypersensitivity to tuberculin among medical students (n=441), nursing students (n=178) and economics students (n=230) in Vitória, State of Espírito Santo, had the aim of assessing whether the prevalence of positive tuberculin tests among healthcare students could be used as a sentinel event for nosocomial transmission in localities with high endemicity. There was a significant change in the frequency of purified protein derivative reactivity in the nursing students (20. 3%) and medical students (18. 4%), in comparison with the economics students (6%) (p <0. 001). As expected, the medical and nursing students had more knowledge about tuberculosis than did the economics students (p <0. 001). Our data suggest that the incidence of Mycobacterium tuberculosis infection among nursing and medical students in our region, which has high endemicity of tuberculosis, does not differ from what is found in low-incidence countries. These data reinforce the need for implementing biosafety programs for this target population.

Lab on paper: biossensores colorimétricos de papel para a detecção de membros do complexo Mycobacterium tuberculosis

Este trabalho foi realizado no âmbito do projecto Lab on Paper, desenvolvido no Centro de Investigação de Materiais (CENIMAT) da Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa (FCT - UNL)

Análise de receptores de quimiocinas na superfície de leucócitos circulantes de indivíduos infectados pelo Mycobacterium leprae: resultados preliminares

Neste estudo, a expressão de receptores de quimiocinas na superfície dos leucócitos circulantes foi feita pela citometria de fluxo. Houve aumento da porcentagem de linfócitos CCR2+CD4+ no sangue periférico dos pacientes com hanseníase. Este resultado preliminar sugeriu alteração do perfil dos receptores de quimiocinas desses pacientes.

Infeção por Mycobacterium Tuberculosis em profissionais de saúde : metodologia de avaliação do risco e sua aplicação num hospital central

RESUMO - A Tuberculose surge, de acordo com o último relatório da Organização Mundial da Saúde, como a segunda principal causa de morte em todo o mundo, de entre as doenças infeciosas. Em 2012, 1.3 milhões de pessoas morreram devido a esta patologia e surgiram 8.6 milhões de novos casos. De entre os grupos de risco de infeção, surgem os profissionais de saúde. A dificuldade no diagnóstico da Tuberculose, o contacto próximo com os pacientes, as medidas de controlo de infeção por vezes inadequadas são algumas das razões que explicam o risco mais elevado de contrair Tuberculose no local de trabalho. Esta Dissertação de Mestrado pretende estabelecer uma nova classificação de risco de infeção por M. tuberculosis em estabelecimentos de saúde, com vista a promover a saúde destes profissionais, inovadora nos critérios de avaliação das medidas de controlo de infeção e de análise dos casos de exposição não protegida a Tuberculose ativa. Esta metodologia de avaliação foi o resultado de uma revisão bibliográfica sobre a temática, tendo sido aplicada num hospital para verificar a sua adequabilidade e mais-valia.

Drug resistance of Mycobacterium tuberculosis strains in southern Brazil

INTRODUCTION: The aim of this work was to evaluate the prevalence of Mycobacterium tuberculosis (MT) strains with mutations that could result in resistance to the main drugs used in treatment in a region with one of the highest numbers of tuberculosis (TB) cases in southern Brazil. METHODS: Deoxyribonucleic acid (DNA) from 120 sputum samples from different patients suspicious of pulmonary tuberculosis who attended the Municipal Public Laboratory for Mycobacterium sp. diagnosis was directly amplified and analyzed by PCR-SSCP. The DNA was amplified in known hotspot mutation regions of the genes rpoB, ahpC, embB, katG, inhA, and pncA. RESULTS: The percentage of samples positive by culture was 9.2% (11/120); 5% (6/120) were positive by bacilloscopy and MT-PCR, and DNA fragments of the aforementioned resistance genes could be amplified from seven (7) of the eleven (11) samples with positive results, either by culture or PCR/bacilloscopy. All presented a SSCP pattern similar to a native, nonresistant genotype, with the ATCC strain 25177 as control, except for one sample (0.01%), which presented a SSCP profile demonstrating mutation at the embB gene. CONCLUSIONS: These results are consistent with the empirical observations by physicians treating TB patients in our region of a low occurrence of cases that are refractory to conventional treatment schemes, in contrast to other parts of the country. Continued surveillance, especially molecular, is essential to detect and monitor the outbreak of MT-resistant strains.

Mediastinitis and pericardial effusion in a patient with AIDS and disseminated Mycobacterium avium infection: a case report

We report the case of a 36-year-old man who had acquired immune deficiency syndrome and developed suppurative mediastinitis extending over the left lung and anterior thoracic wall around the sternum, pericardial effusions, splenomegaly, and mesenteric and periaortic lymphadenomegaly due to Mycobacterium avium (genotype I). The organism was isolated from an axillary lymph node and the bone marrow. Mediastinitis associated with disseminated M. avium complex infection is uncommon and, to the best of our knowledge, this manifestation has not reported before.

Mycobacterium fortuitum-related lymphadenitis associated with the varicella-zoster virus

Lymphadenitis caused by non-tuberculous mycobacteria is an uncommon manifestation in immunocompetent individuals. Here, we report a case of Mycobacterium fortuitum infection in a previously healthy 9-year-old patient who developed cervical lymphadenitis evolving to a suppurative ulcer associated with a varicella-zoster virus infection. We discuss the relationship between the varicella-zoster virus and the immune response of the host as an explanation for the unusual progression of the case.

Prevalence and evolution of Mycobacterium tuberculosis infection in tuberculosis case contacts

INTRODUCTION : The tuberculin test is a diagnostic method for detecting latent tuberculosis (TB) infection, especially among disease contact cases. The objective of this study was to analyze the prevalence and evolution of Mycobacterium tuberculosis infection among TB contact cases. METHODS : A retrospective cohort study was performed in a reference center for TB. The study population consisted of 2,425 patients who underwent a tuberculin test from 2003 to 2010 and whose results indicated contact with individuals with TB. The data were collected from the registry book of the tuberculin tests, patient files and the Information System Records of Notification Grievance. To verify the evolution of TB, case records through September 2014 were consulted. Data were analyzed using the Statistical Package for the Social Sciences (SPSS). In all hypothesis tests, a significance level of 0.05 was used. RESULTS : From the studied sample, 435 (17.9%) contacts did not return for reading. Among the 1,990 contacts that completed the test, the prevalence of latent TB infection was 35.4%. Of these positive cases, 50.6% were referred to treatment; the dropout rate was 42.5%. Among all of the contacts, the TB prevalence was 1.8%, from which 13.2% abandoned treatment. CONCLUSIONS : The collected data indicate the need for more effective public policies to improve TB control, including administering tests that do not require a return visit for reading, enhancing contact tracing and encouraging actions that reinforce full treatment adherence.

Diagnosis of latent Mycobacterium tuberculosis infection: tuberculin test versus interferon-gamma release

Abstract: INTRODUCTION: The treatment of individuals with active tuberculosis (TB) and the identification and treatment of latent tuberculosis infection (LTBI) contacts are the two most important strategies for the control of TB. The objective of this study was compare the performance of tuberculin skin testing (TST) with QuantiFERON-TB Gold In TUBE(r) in the diagnosis of LTBI in contacts of patients with active TB. METHODS: Cross-sectional analytical study with 60 contacts of patients with active pulmonary TB. A blood sample of each contact was taken for interferon-gamma release assay (IGRA) and subsequently performed the TST. A receiver operating characteristic curve was generated to assess the cutoff points and the sensitivity, predictive values, and accuracy were calculated. The agreement between IGRA and TST results was evaluated by Kappa coefficient. RESULTS: Here, 67.9% sensitivity, 84.4% specificity, 79.1% PPV, 75% NPV, and 76.7% accuracy were observed for the 5mm cutoff point. The prevalence of LTBI determined by TST and IGRA was 40% and 46.7%, respectively. CONCLUSIONS: Both QuantiFERON-TB Gold In TUBE(r) and TST showed good performance in LTBI diagnosis. The creation of specific diagnostic methods is necessary for the diagnosis of LTBI with higher sensitivity and specificity, preferably with low cost and not require a return visit for reading because with early treatment of latent forms can prevent active TB.

Single-tube nested PCR assay with in-house DNA extraction for Mycobacterium tuberculosis detection in blood and urine

Abstract: INTRODUCTION : Molecular analyses are auxiliary tools for detecting Koch's bacilli in clinical specimens from patients with suspected tuberculosis (TB). However, there are still no efficient diagnostic tests that combine high sensitivity and specificity and yield rapid results in the detection of TB. This study evaluated single-tube nested polymerase chain reaction (STNPCR) as a molecular diagnostic test with low risk of cross contamination for detecting Mycobacterium tuberculosis in clinical samples. METHODS: Mycobacterium tuberculosis deoxyribonucleic acid (DNA) was detected in blood and urine samples by STNPCR followed by agarose gel electrophoresis. In this system, reaction tubes were not opened between the two stages of PCR (simple and nested). RESULTS: STNPCR demonstrated good accuracy in clinical samples with no cross contamination between microtubes. Sensitivity in blood and urine, analyzed in parallel, was 35%-62% for pulmonary and 41%-72% for extrapulmonary TB. The specificity of STNPCR was 100% in most analyses, depending on the type of clinical sample (blood or urine) and clinical form of disease (pulmonary or extrapulmonary). CONCLUSIONS: STNPCR was effective in detecting TB, especially the extrapulmonary form for which sensitivity was higher, and had the advantage of less invasive sample collection from patients for whom a spontaneous sputum sample was unavailable. With low risk of cross contamination, the STNPCR can be used as an adjunct to conventional methods for diagnosing TB.

Molecular characterization of Mycobacterium tuberculosis isolates from Tehran, Iran by restriction fragment length polymorphism analysis and spoligotyping

Abstract: INTRODUCTION Characterization of Mycobacterium tuberculosis (MTB) isolates by DNA fingerprinting has contributed to tuberculosis (TB) control. The aim of this study was to determine the genetic diversity of MTB isolates from Tehran province in Iran. METHODS MTB isolates from 60 Iranian and 10 Afghan TB patients were fingerprinted by standard IS6110-restriction fragment length polymorphism (RFLP) analysis and spoligotyping. RESULTS The copy number of IS6110 ranged from 10-24 per isolate. The isolates were classified into 22 clusters showing ≥ 80% similarity by RFLP analysis. Fourteen multidrug-resistant (MDR) isolates were grouped into 4 IS6110-RFLP clusters, with 10 isolates [71% (95% CI: 45-89%)] in 1 cluster, suggesting a possible epidemiological linkage. Eighteen Iranian isolates showed ≥ 80% similarity with Afghan isolates. There were no strains with identical fingerprints. Spoligotyping of 70 isolates produced 23 distinct patterns. Sixty (85.7%) isolates were grouped into 13 clusters, while the remaining 10 isolates (14.2%) were not clustered. Ural (formerly Haarlem4) (n = 22, 31.4%) was the most common family followed by Central Asian strain (CAS) (n = 18, 25.7%) and T (n = 9, 12.8%) families. Only 1strain was characterized as having the Beijing genotype. Among 60 Iranian and 10 Afghan MTB isolates, 25% (95% CI: 16-37) and 70% (95% CI: 39-89) were categorized as Ural lineage, respectively. CONCLUSIONS A higher prevalence of Ural family MTB isolates among Afghan patients than among Iranian patients suggests the possible transmission of this lineage following the immigration of Afghans to Iran.

Molecular tools in the diagnostic and epidemiology of infections caused by members of Mycobacterium avium Complex

Mycobacterium avium Complex (MAC) comprises microorganisms that affect a wide range of animals including humans. The most relevant are Mycobacterium avium subspecies hominissuis (Mah) with a high impact on public health affecting mainly immunocompromised individuals and Mycobacterium avium subspecies paratuberculosis (Map) causing paratuberculosis in animals with a high economic impact worldwide. In this work, we characterized 28 human and 67 porcine Mah isolates and evaluated the relationship among them by Multiple-Locus Variable number tandem repeat Analysis (MLVA). We concluded that Mah population presented a high genetic diversity and no correlations were inferred based on geographical origin, host or biological sample. For the first time in Portugal Map strains, from asymptomatic bovine faecal samples were isolated highlighting the need of more reliable and rapid diagnostic methods for Map direct detection. Therefore, we developed an IS900 nested real time PCR with high sensitivity and specificity associated with optimized DNA extraction methodologies for faecal and milk samples. We detected 83% of 155 faecal samples from goats, cattle and sheep, and 26% of 98 milk samples from cattle, positive for Map IS900 nested real time PCR. A novel SNPs (single nucleotide polymorphisms) assay to Map characterization based on a Whole Genome Sequencing analysis was developed to elucidate the genetic relationship between strains. Based on sequential detection of 14 SNPs and on a decision tree we were able to differentiate 14 phylogenetic groups with a higher discriminatory power compared to other typing methods. A pigmented Map strain was isolated and characterized evidencing for the first time to our knowledge the existence of pigmented Type C strains. With this work, we intended to improve the ante mortem direct molecular detection of Map, to conscientiously aware for the existence of Map animal infections widespread in Portugal and to contribute to the improvement of Map and Mah epidemiological studies.

Pesquisa de Micobactérias Ambientais em água de torneira, luvas e soluções utilizadas em procedimentos cirúrgicos no Hospital Universitário Getúlio Vargas - Manaus/AM

Investigou-se por métodos bacteriológicos (cultivo) e moleculares (PCR - Restriction Enzyme Analysis, PRA), a presença de micobactérias ambientais em águas de torneira, soluções e luvas cirúrgicas, utilizadas nas etapas dos procedimentos cirúrgicos executados no centro cirúrgico do Hospital Universitário Getulio Vargas (HUGV), na cidade de Manaus-Amazonas/Brasil. Foram colhidas e analisadas 105 amostras sendo: 24 de águas (colhidas das 2 torneiras existentes no centro cirúrgico), 8 de solução de Povidine e 7 de solução de Clorhexidina, que servem para a higienização das mãos dos cirurgiões; 39 de luvas cirúrgicas (superfícies internas e externas); e 27 de soluções que foram efetivamente utilizadas durante o ato cirúrgico. Por método bacteriológico obteve-se 41 isolados micobacterianos apenas de águas das torneiras. Pelo PRA obteve-se a detecção de DNA micobacteriano somente na amostra de água que forneceu acima de 100 colônias de micobactérias por tubo semeado. Os isolados foram identificados como sendo Mycobacterium celatum perfil 2, M. gordonae perfil 3, M. gordonae perfil 6, M. intracellulare perfil 1, M. lentiflavum perfil 3 e M. mucogenicum perfil 1. O encontro de M. mucogenicum, espécie já incriminada em surtos pós-cirúrgicos, indica que devem ser efetuados procedimentos de limpeza e monitoramento em todos os pontos de distribuição de águas, visando à prevenção de surtos de micobacterioses nosocomiais induzidos pelo uso das águas nas diferentes atividades de manuseio ou higienização dos pacientes submetidos a procedimentos invasivos.