Tuberculosis osteoarticular en un paciente pediátrico / Osteoarticular tuberculosis in a pediatric patient

Introducción: La tuberculosis osteoarticular es una enfermedad inflamatoria crónica, muy rara con un cuadro clínico atípico y se presenta con una incidencia de 1-2 por ciento del total de los casos de tuberculosis. Objetivo: Exponer una forma de presentación poco frecuente de tuberculosis en Honduras Presentación del caso: Se trata de una niña de dos años con diagnóstico inicial de sinovitis en rodilla derecha de cinco meses de evolución y tratada con antibiótico y analgésico. Sin mejoría se presenta a emergencia con cambios inflamatorios. Se realiza rayos x de rodilla que muestran lesiones osteolíticas a nivel de rótula y cóndilo del fémur derecho. Baciloscopia de esputo y prueba de tuberculina negativas. Familiar de tercer grado positivo para tuberculosis hacía un año y medio. Por biopsia de tejido blando y óseo de rodilla derecha se establece el diagnóstico de artritis por Mycobacterium tuberculosis por estudio inmuno-histoquimico con tinción Ziehl Nielsen. Conclusiones: Es el primer informe de caso de tuberculosis osteoarticular en un paciente pediátrico descrito en Honduras. Por la larga evolución de la enfermedad, lo atípico de su clínica y su baja incidencia es difícil establecer el diagnóstico final. Fue imprescindible el estudio anatomopatológico por biopsia que permitiera esclarecer a los clínicos el diagnóstico e iniciar el tratamiento oportuno(AU)

Introduction: Osteoarticular tuberculosis is a chronic inflammatory disease, very rare, and with an atypical clinical picture and occurs with an incidence of 1-2 percent of all TB cases. Objective: Show a rare form of TB´s presentation in Honduras Case presentation: Two-year-old girl with an initial diagnosis of right knee synovitis of five months of evolution and treated with antibiotics and analgesics. Without improvement, she attends to emergencies service with inflammatory changes. Knee x-rays show osteolytic lesions at the kneecap level and the condyle of the right femur. Sputum bacilloscopy and negative tuberculin test were performed. She had a third-grade relative positive to tuberculosis a year and a half ago. A right knee soft tissue and bone biopsies confirm the diagnosis of arthritis by Mycobacterium tuberculosis by immuno-histochemical study with Ziehl Nielsen staining. Conclusions: It is the first osteoarticular TB case report in a pediatric patient described in Honduras. Because of the long evolution of the disease, the atypicalness of its clinic features and its low incidence, it is difficult to establish the final diagnosis. Anatomopathological study by biopsy was essential to clarify the diagnosis to clinicians and initiate timely treatment(AU)

Exposure to cetyl pyridinium chloride and loss of integrity of cell wall of mycobacteria.

Background: Cetyl pyridinium chloride (CPC) liquefied sputum was shown to reduce AFB smear positivity presumably damaging cell wall of M. tuberculosis. Settings: National Institute for Research in Tuberculosis, Chennai, (Tamil Nadu). Objective: To assess the cell wall damage of mycobacteria in CPC liquefied sputum, by Transmission Electron Microscopy (TEM) and mycobacteriophage adsorption studies. Methods: Pooled sputum sample from smear positive pulmonary TB patients was homogenized and liquefied with CPC. It was examined in TEM daily for four days, to assess cell wall damage of M. tuberculosis, and photomicrographs were taken. M. smegmatis mc2155, treated with CPC, was infected with mycobacteriophage (phAE129) to study phage adsorption on cell wall and plaque formation. CPC untreated sputum and M. smegmatis formed controls. Results: Photomicrographs showed that cell wall of M. tuberculosis was intact in controls and damaged in CPC preserved sputum for 96 hours. Plaque formation was seen and absent respectively in CPC untreated and treated M. smegmatis cells. Conclusion: Exposure to CPC damaged the cell wall of M. tuberculosis within 96 hours. Mycobacteriophage failed to form plaques after M. smegmatis mc2155 was treated with CPC implying inhibition of phage adsorption on damaged cell wall and thus providing a clue for poor staining and smear positivity in microscopy.

Mycobacterium under AFM tip: advantages of polyelectrolyte modified substrate

Immobilization of bacterial cells onto surfaces is critical for imaging with an atomic force microscope. In this paper, polyethylenimine [PEI] coated silicon plates are shown to be suitable for immobilizing and imaging Mycobacterium tuberculosis

Detección de mutaciones en los Genes rpoß y katG asociados a resistencia en cepas de Mycobacterium tuberculosis de Bolivia / Detection of mutations in rpoß and katG genes associated to resistance in Mycobacterium tuberculosis strains from Bolivia

Objetivo: Evaluar la sensibilidad y especificidad de la genotipificación, como instrumento de diagnóstico rápido y confiable parala detección de mutaciones en los genes rpoß y katG asociados a resistencia,en cepas de Mycobacterium tuberculosis de Bolivia.Diseño: Test Diagnóstico Metodología: Las cepas analizadas fueron aisladas y enviadas por los diferentes Laboratorios de la Red Nacional de Diagnósticode Tuberculosis de Bolivia entre febrero y diciembre de 2007. La muestra para el presente estudio estuvo constituida por un totalde 65 aislamientos previamente caracterizados por métodos fenotípicos de cultivo y pruebas de sensibilidad a la RIF e INH, porel método de las proporciones Canetti-Rist. La genotipificación ha sido realizada utilizando el kit Genotype MTBDR, basado enla utilización de métodos de amplificación e hibridización, para detectar mutaciones a nivel de los marcadores de resistencia rpoßy katG.Resultados: Se procedió al cálculo de la sensibilidad y especificidad de la prueba de diagnóstico; además de los valores predictivospositivo y negativo. Dicho análisis muestra los siguientes resultados: sensibilidad 74...

Objective:To evaluate the sensitivity and specificity of genotyping as a tool for rapid and reliable detection of mutations in rpoß and katG genes associated with resistance in Mycobacterium tuberculosisstrains from Bolivia. Design:Diagnostic Test Methodology: The strains analyzed were isolated and submitted by different laboratories of the National Network for Diagnosisof Tuberculosis of Bolivia between February and December 2007. The sample for this study consisted of 65 isolates previousl y characterized by phenotypic methods of culture and sensitivity testing to RIF and INH by the Canetti-Rist proportion method. Genotyping of these samples has been done using the MTBDR Genotype kit, according to amplification and hybridization methodsto detect mutations at the rpoß and katG resistance markers.Results:The sensitivity and specificity of the diagnostic tests were calculated, as well as the positive and negative predictivevalues. This analysis shows the following results: sensitivity 74%, specificity 92%, positive predictive value 92%, and negative predictive value 73%. Conclusions: The genotyping test using Genotype MTBDR, meeting validation criteria for a diagnostic test study in our country,constitutes a quick, useful and reliable tool for use in diagnosis and routine determination of sensitivity and resistance in MTBCstrains.

Nuevos métodos para el diagnóstico de la tuberculosis / New methods for the diagnosis of tuberculosis

La magnitud del problema mundial de la tuberculosis y su potencial de incremento han conducido a la necesidad de mejorar los métodos de diagnóstico como una de las estrategias conducentes al control de la enfermedad. Los nuevos métodos incluyen pruebas moleculares para la detección directa, medios de cultivo rápido y novedosos procedimientos de identificación. Para la detección directa de micobacterias se dispone hoy de pruebas basadas en la amplificación de los ácidos nucleicos, cuya sensibilidad supera a la de la baciloscopia y es cercana a la de los cultivos, pero con mayores costos que los procedimientos tradicionales.La automatización para detectar el crecimiento en medios de cultivo produjo un cambio radical en la micobacteriología, pues con ella se logró disminuir considerablemente el tiempo requerido para aislar el M. tuberculosis (MTB); la combinación de métodos de detección rápida permite en la actualidad mejores tasas de aislamiento y la demostración más precoz del crecimiento. Las nuevas alternativas han logrado aumentar tanto la sensibilidad como la especificidad y la rapidez diagnósticas; sin embargo, es necesario mejorar aún más los métodos de diagnóstico de esta enfermedad de modo que se combinen el mejor desempeño y la simplicidad de los procedimientos con la disminución de los costos.

The magnitude of the global tuberculosis problem and its potential for increase have led to efforts to improve the diagnostic methods as one of the leadingcontrol strategies. New diagnostic tools have been developed, including direct detection tests, rapid culture media and identification methods. For the direct detection of mycobacteria, tests based on nucleic acid amplification have been implemented, with higher sensitivity than the direct smear, close to that of the cultures; however, they are still expensive methods. Availability of the automated cultures resulted in a radical change in mycobacteriology, reducing the time required for the isolation ofMycobacterium tuberculosis. The combination of rapid methods has increased the rates of isolation and decreased the time required for the detection of growth. Different tools for identifying mycobacteria have also been developed, aimed at improving theperformance of traditional biochemical methods and achieving species identification in a few hours. The new diagnostic alternatives have succeeded in improving the sensitivity, specificity and speed in the diagnosis of tuberculosis; however, the challengeremains to reduce their cost and to make them more accessible in countries where the problem with this disease is still serious.

Auramine phenol staining of smears for screening acid fast bacilli in clinical specimens.

Tuberculosis is a major threat to the public health allover world. Out of the total tuberculosis cases reported globally, more than half are reported from African continent and India. Two to three fold rises in tuberculosis cases has been reported in the last two decades. Early diagnosis and treatment is one of the effective tools to control the rapid spread of disease. The aim of this study was to find out the value of auramine phenol (AP) staining technique in diagnosis of the suspected tuberculosis cases. A total of 2000 samples which included sputum (746), gastric aspirates (380), urine (336), endometrial biopsy (150), pleural fulids (146), Synovial fluids (67), ascitic fluids (35), cerebrospinal fluids (43), bone marrow (18), lymph node biopsy (11), pericardial aspirates (6), skin biopsy (4), peritoneal fluids (2), and stool (1) were included in the study. Sample were subjected for decontamination procedure by using standard Petroffs method. The deposit smears were stained by auramine phenol (AP stain) and Ziehl-Neelsenstaining (ZN stain) and specimens were cultured for Mycobacterium tuberculosis. Of the total positive isolates 69.23% were having pulmonary tuberculosis and 30.76 had extrapulmonary tuberculosis Genitourinary tuberculosis was the most common diagnosis among the extrapulmonary tuberculosis followed by chronic synovitis, bursitis, meningitis, septic arthritis and pericardial effusion. Out of 130 positive samples 70 by culture, 66 smears were positive by auramine phenol stain and 62 were positive by ZN stain. A total of 27 samples were tested positive only by AP staining technique, which included (12) pulmonary and (15) extrpulmonary samples. The endometrial biopsy and pericardial fluid samples showed positive for acid fast bacilli by AP stain only, whereas ZN stain and culture technique failed to demonstrate any bacilli in the same sample. Auramine stain showed high sensitivity (47.14%) and specificity (96.58%). Result of the present study showed that the auramine stain is a better method for screening samples from the suspected cases of tuberculosis sample especially pulmonary and extrapulmonary cases where bacilli count is usually low.

[Morphologic changes of Mycobacterium tuberculosis after exposure with choloformic garlic extract]

Tuberculosis is one of the major problem facing of globle health. Drug resistance of mycobacterium tuberculosis to antimicrobial agent has strongly emerged the need for achiving the new drugs. Garlic as medical plants has long been taken under investigation. This study for antibacterial effect was done to determine the morphological alteration of Mycobacterium tuberculosis due to garlic choloformic extract. Garlic extract contains allicine [thio-2-propen-sulfonic acid-s-allil ester] is one of its effective antimicrobacterial substance. In a in-vitro study, the standard strain of Mycobacterium tuberculosis H37RV and clinical isolated strain was cultured in the middle broke 7H9 broth with different concentration of garlic extract in different 12, 24, 48, 72 hours. Morphological althertits of mycobacterium inspected with macroscopic and microscopic studies. The garlic exteract caused conversion of rough colonies to smooth and mucoid colonies and in microscopic studies morphologic change of mycobacterium from bacilli form to coccobacilli and cocci was observed. Also 0.67 mg/ml of garlic exteract on 48h period inhibited both of sensitive [standard strain of H37RV] and resistance [clinical strains] Mycobacterium tuberculosis. This study showed that garlic extract in addition to inhibiting growth, change the morphology of Mycobacterium tuberculosis from baccilli to cocoibaccill form and also alter the colony apearance from rough to smooth shap

Avaliaçao de um novo sistema para detecçao de micobactérias: "mycobacterium growth indicator tube" (MIGIT) / Evaluation of a new system for the detection of mycobacteria: mycobacterium grouth indicator tube (MGIT)

Com o objetivo de avaliar uma nova metodologia para detectar o crescimento de micobactérias, analisamos 159 materiais clínicos previamente selecionados pelo método de Ziehl-Neelsen (149 positivos e 10 negativos), provenientes de diversos pacientes do Hospital das Clínicas da FMUSP. As amostras foram descontaminadas e concentradas pelo método do NALC (N-acetil-L-cisteína), examinadas pela coloraçao de Ziehl-Neelsen e 0,5ml de cada amostra tratada foi inoculada diretamente no MGIT ("Mycobacterium Growth Indicator Tube", Becton Dickinson Microbiology Systems, EUA) e o mesmo volume foi inoculado no meio e Lowenstein-Jensen (LJ). Os tubos de MIGT e de LJ foram incubados a 37 graus e observados diariamnte quanto ao crescimento e emissao de fluorescência. Para as leituras do MGIT foi utilizada uma luz UV de 365 nm e as culturas no meio de LJ foram observadas visualmete quanto ao crescimento. O MGIT contém 4,5 ml de meio Middelbrook 7H9 enriquecido e um sensor de fluorecência na base do tubo, sensíve ao oxigênio. Havendo crescimento de micobactérias, o seu metabolismo resulta em um consumo de oxigênio, ocasionando um aumento de fluorescência. Das 149 amostras com pesquisa positiva pelo método de ZIehl-Neelsen, 144 amostras foram positivas no MGIT, sendo 53 positivas na primeira semana e 78 na segunda semana de incubaçao. Das 131 amostras positivas no meio de LJ, a maioria apresentou crecimento após 15 dias de incubaçao. O MIGIT provou ser um sistema simples, rápido e sensível para detectar o crecimento de micobactérias diretamente de espécimes clínicos

Acción del Agua del Volcán Copahue (Neuquén, Argentina) sobre la morfología de las microbacterias / Action of the Copahue Volcano (Neuquén, Argentine) water of mycobateria morphology

Se efectuó un estudio de la acción del Agua del Volcán Copahue (AVC), Neuquén, Argentina, sobre 15 cepas de microbacterias Mycobacterium tuberculosis: M. bovis y los mycobacterum no tuberculosos ("Atipicos"), poniendo especial interés en los que forman "cuerdas". Se utilizó AVC con su pH l,3 y se la llevó a pH 6,5. En los bacilos que resistieron la acción del agua hasta el momento de la coloración, se advirtieron elementos más o menos alterados. Al aumentar el tiempo de contacto se llegó a la destrucción total, observándose en algunos casos muy pocos bacilos aislados, material deteriorado y formas granulares. Estas alteraciones fueron mucho más marcadas en las suspensiones que en el líquido del sedimento con el AVC; en las primeras directamente no se llegó a reparar la formación de cuerdas en ningun momento aun en bacilos que deberían formarla. Su acción no estaría asociada a una reacción bioquímica responsable en la síntesis de la pared celular, como la transpeptidación. El AVC actuaría sobre la síntesis del ácido micólico y se trataría de un agente bacteriolítico. Además se realizaron estudios en el "Laboratorio de tratamiento de imágenes", INEUCI (Instituto de Neurociencia), CONICET