Neumonía lipoidea exógena crónica y sobreinfección por Mycobacterium tuberculosis / Chronic Exogenous Lipoid Pneumonia and Superinfection by Mycobacterium Tuberculosis

Introducción: la neumonía lipoidea exógena es una enfermedad pulmonar inflamatoria poco común, desencadenada por la inhalación o aspiración de material graso de origen animal, vegetal o mineral. El diagnóstico se establece a través de confirmación histo-patológica, por la presencia de macrófagos cargados de lípidos en muestras respirato-rias, asociado a las características clínicas específicas al momento de su presentación.Requiere de un alto nivel de sospecha y una adecuada anamnesis de los antecedentes exposicionales del paciente debido a que muchos casos son subdiagnosticados y trat-ados como neumonía adquirida en la comunidad, lo que retrasa su diagnóstico y mane-jo, sumado a la ausencia de guías disponibles para su tratamiento.Se han reportado pocos casos de sobreinfección por tuberculosis en pacientes con neu-monía lipoidea exógena crónica. Caso clínico: femenino 33 años, con antecedentes de exposición crónica a sustancias desinfectantes de características aceitosas sin protección de vía aérea, con cuadro de tos y dolor torácico. Conclusión: el diagnóstico temprano, asociado a tratamiento de soporte, general-mente conservador, favorece la mejoría clínica y radiológica, y de esta manera dis-minuye la morbimortalidad. (AU)

Introduction: exogenous lipoid pneumonia is a rare inflammatory lung disease, trigge-red by inhalation or aspiration of fatty material of animal, vegetable or mineral origin. The diagnosis is established through histological confirmation by the presence of lipid-laden macrophages in respiratory samples, associated with the specific clinical charac-teristics at the time of presentation. It requires a high level of suspicion and an adequate anamnesis of the patient's expo-sure history, since many cases are underdiagnosed and treated as community-acquired pneumonia, what delays its diagnosis and management, added to the absence of avai-lable guidelines for its treatment. Few cases of tuberculosis superinfection have been reported in patients with exoge-nous lipoid pneumonia.Clinical case: 33-year-old female, with a history of chronic exposure to oily disinfectant substances without airway protection, with symptoms of cough and chest pain.Conclusion: early diagnosis, associated with supportive treatment, generally conser-vative, favors clinical and radiological improvement, thus reducing morbidity and mor-tality. (AU)

Aplicación de PCR a partir de cultivo en la identificación de subespecies del complejo Mycobacterium Tuberculosis / The use of culture-based PCR for the identification of subspecies of the mycobacterium tuberculosis complex

Los métodos diagnósticos clásicos de tuberculosis (TB) se basan en la utilización de baciloscopía y cultivo. La identificación del agente etiológico desde la positivización del cultivo requiere entre 10 y 15 días, mientras que el empleo de la reacción en cadena de la polimerasa (PCR) disminuye el tiempo a 24 h, lo que permite no solo identificar las subespecies del complejo Mycobacterium tuberculosis (CMTB) sino también diferenciarlas de otras especies ambientales clínicamente importantes (MOTT) facilitando el diagnóstico y tratamiento. El objetivo del presente trabajo fue determinar la utilidad de la PCR en la identificación temprana de las micobacterias pertenecientes al CMTB, a partir de cultivos positivos, de pacientes con sospecha de TB, atendidos en un hospital pediátrico de alta complejidad, durante un período de cuatro años. A cada muestra, se le realizó baciloscopía y cultivo en medio líquido. A los cultivos positivos, una inmunocromatografía lateral (TBIDR) y luego PCR. El 4,6% del total de muestras (510/11.162) pertenecientes a 198 pacientes presentó cultivos positivos. Cuatrocientos veintiseis (84%) correspondieron a muestras respiratorias. El rendimiento de la baciloscopía directa fue del 41% (194/470). Cuatrocientos treinta y ocho (86%) resultaron M. tuberculosis, 21 (4%) Mycobacterium bovis, 7 (1%), M. bovis-BCG y 44 (9%) MOTT. La utilización de medios de cultivos líquidos junto con el empleo de PCR favorecen una rápida orientación microbiológica y constituye una estrategia útil para optimizar el manejo clínico de estas infecciones, desde el punto de vista terapéutico y epidemiológico, especialmente en pediatría (AU)

Classical diagnostic methods for tuberculosis (TB) are based on the use of smear microscopy and culture. The identification of the etiological agent from positive culture requires 10 to 15 days, while the use of the polymerase chain reaction (PCR) reduces the time to 24 h, which allows not only to identify the subspecies of the Mycobacterium tuberculosis complex (MTC) but also to differentiate them from clinically important environmental mycobacteria other than tuberculosis (MOTT), facilitating diagnosis and treatment. The aim of this study was to determine the usefulness of PCR in the early identification of mycobacteria belonging to the MTC, from positive cultures of patients with suspected TB seen in a pediatric tertiary hospital over a 4-year period. For each sample, smear microscopy and culture in liquid medium was performed. Positive cultures were subjected to lateral immunochromatography (TBIDR) and then PCR. Of the total number of samples (510/11,162) belonging to 198 patients, 4.6% showed positive cultures; 426 (84%) were respiratory samples. The direct smear microscopy yield was 41% (194/470). Overall, 438 (86%) were found to be M. tuberculosis, 21 (4%) Mycobacterium bovis, 7 (1%), M. bovis-BCG, and 44 (9%) MOTT. The use of liquid culture media together with the use of PCR favors a rapid microbiological orientation and is a useful strategy to optimize the clinical management of these infections, from a therapeutic and epidemiological point of view, especially in children (AU)

Comparison of different diagnostic modalities for isolation of Mycobacterium Tuberculosis among suspected tuberculous lymphadenitis patients / Comparação de diferentes modalidades de diagnóstico para isolamento de Mycobacterium tuberculosis entre pacientes com suspeita de linfadenite tuberculosa

Abstract Tuberculosis is a communicable disease with high morbidity and mortality rates in developing countries. The study's primary objective is to compare conventional methods such as acid-fast bacillus (AFB) culture and microscopy with rapid diagnostic methods. The secondary objective is to compare histopathological and microbiological findings in suspected patients with tubercular lymphadenitis. A total of 111 samples (August 2018 to September 2019) of lymph nodes were processed for AFB microscopy, AFB cultures, drug-susceptibility testing (DST), histopathology, and Xpert Mycobacterium Tuberculosis (MTB)/resistance to Rifampin (RIF) assays. Out of 111 lymph node samples, 6 (5.4%) were positive for AFB smear microscopy, 84 (75.6%) were positive for AFB culture, 80 (70.7%) were positive on Gene Xpert, and 102 (91.8%) were indicative of tuberculosis for histopathology studies. Mycobacteria growth indicator tube (MGIT) culture positivity was 84 (75.6%) higher than solid Lowenstein-Jensen (LJ) culture 74 (66.6%). Positive cultures underwent phenotypic DST. Two cases were Multidrug-resistant (MDR) on DST, while three cases were Rifampicin resistant on Gene Xpert. The sensitivity of Genexpert was (62%) against the conventional AFB culture method. The poor performance of conventional lymphadenitis diagnostic methods requires early and accurate diagnostic methodology. Xpert MTB/RIF test can help in the treatment of multidrug-resistant TB cases. Nonetheless, rapid and conventional methods should be used for complete isolation of Mycobacterium tuberculosis.

Resumo A tuberculose é uma doença transmissível com altas taxas de morbimortalidade nos países em desenvolvimento. O objetivo principal do estudo é comparar métodos convencionais, como cultura de bacilo álcool-ácido resistente (BAAR) e microscopia, com métodos de diagnóstico rápido. O objetivo secundário é comparar os achados histopatológicos e microbiológicos em pacientes com suspeita de linfadenite tubercular. Um total de 111 amostras (agosto de 2018 a setembro de 2019) de gânglios linfáticos foi processado ​​para microscopia de AFB, culturas de AFB, teste de susceptibilidade a drogas (DST), histopatologia e Xpert Mycobacterium tuberculosis (MTB)/ensaios de resistência à rifampicina (RIF). Das 111 amostras de linfonodos, 6 (5,4%) foram positivas para baciloscopia de AFB, 84 (75,6%) foram positivas para cultura de AFB, 80 (70,7%) foram positivas para o GeneXpert e 102 (91,8%) foram indicativas de tuberculose para estudos histopatológicos. A positividade da cultura do tubo indicador de crescimento de micobactérias (MGIT) foi 84 (75,6%), maior que a cultura sólida de Lowenstein-Jensen (LJ), 74 (66,6%). As culturas positivas foram submetidas a DST fenotípico. Dois casos eram multirresistentes (MDR) ao DST, enquanto três casos eram resistentes à rifampicina no GeneXpert. A sensibilidade do GeneXpert foi 62% contra o método convencional de cultura AFB. O fraco desempenho dos métodos convencionais de diagnóstico de linfadenite requer metodologia de diagnóstico precoce e precisa. O teste Xpert MTB/RIF pode ajudar no tratamento de casos de tuberculose multirresistente. No entanto, métodos rápidos e convencionais devem ser usados ​​para o isolamento completo do Mycobacterium tuberculosis.

Vigilancia de la resistencia del Mycobacterium tuberculosis a los fármacos en Paraguay, 2014 a 2017 / Surveillance of drug resistance of Mycobacterium tuberculosis in Paraguay, 2014 to 2017

El objetivo del estudio fue identificar la resistencia del Mycobacterium tuberculosis a los fármacos en Paraguay, 2014 a 2017. Se realizó un estudio observacional retrospectivo. Se utilizaron los datos del Programa Nacional de Tuberculosis del Paraguay comprendidos entre los años 2014 a 2017. Se incluyeron todos los pacientes con diagnóstico de Tuberculosis que se realizaron un test de resistencia. Se extrajeron los datos en Excel y fueron analizados con Stata 17.0. Se incluyeron 3429 pacientes con tuberculosis que contaban con resultado de al menos una prueba de sensibilidad. La resistencia se encontró en 2.1% de los pacientes. La resistencia a la Rifampicina estuvo presente en el 0.3% de los casos mientras que a la Izionazida en el 0.6% de los casos. La prevalencia de resistencia fue más alta en hombres 3.4 (IC 95% 2.2 - 4.8) p=0.003, que residían en el chaco 6.0 (IC 95% 3.4 - 9.7) p=0.000, previamente tratados 2.7 (IC 95% 1.1 - 5.1) p=0.010. En el modelo se pudo observar que un paciente previamente tratado tiene mayores posibilidades de tener resistencia OR 2.62 (IC 95% 1.1 - 6.24). La prevalencia de resistencia del Mycobacterium tuberculosis a fármacos estuvo relacionada con haber sido previamente tratado

The objective of the study was to identify the resistance of Mycobacterium tuberculosis to drugs in Paraguay, 2014 to 2017. A retrospective observational study was carried out. The data from the National Tuberculosis Program of Paraguay between the years 2014 to 2017 were used. All patients with a diagnosis of Tuberculosis who underwent a resistance test were included. Data were extracted in Excel and analyzed with Stata 17.0. 3429 tuberculosis patients who had a result of at least one sensitivity test were included. Resistance was found in 2.1% of patients. Resistance to Rifampicin was present in 0.3% of cases while to Izionazide in 0.6% of cases. The prevalence of resistance was higher in men 3.4 (95% CI 2.2 - 4.8) p = 0.003, who resided in the Chaco 6.0 (95% CI 3.4 - 9.7) p = 0.000, previously treated 2.7 (95% CI 1.1 - 5.1) p = 0.010. In the model, it was observed that a previously treated patient has a greater chance of having resistance OR 2.62 (95% CI 1.1 - 6.24). The prevalence of resistance of Mycobacterium tuberculosis to drugs was related to having been previously treated

Rapid identification of Mycobacterium tuberculosis in cultures by molecular and immunological methods / Identificação rápida de Mycobacterium tuberculosis em culturas por método molecular e método imunológico

Objective: The study aimed to evaluate molecular and immunological methods and to propose a workflow using them for tuberculosis (TB) diagnosis routine. Methods: A cross-sectional retrospective study was performed, including 121 liquid cultures from a TB laboratory located in the extreme south of Brazil. All cultures were positive for Mycobacterium tuberculosis complex (MTBC) by in-house Polymerase Chain Reaction (PCR) using DNA extracted by the CTAB method (PCR-CTAB) for IS6110 detection. These cultures were subjected to faster tests than this one, the immunological MPT64 assay and the PCR using DNA extracted by thermal lysis method (PCR-TL), and these were evaluated for MTBC identification using PCR-CTAB as a reference method. Results: The sensitivity of MPT64 assay and PCR-TL to identify MTBC in positive cultures by PCR-CTAB were 73.6% (89/121) and 98.3% (119/121), respectively. We proposed a workflow based on the use of MPT64 assay in liquid cultures suggestive of MTBC, and in case of a negative result, we suggest the performance of PCR-TL. The PCR-CTAB is suggested only if faster tests are negative. Conclusions: Methods capable of confirming MTBC in cultures should continue to be standardized, tested, and optimized to meet the ideal requirements of simplicity, quickness, and effectiveness. The molecular and immunological methods evaluated have differences in the execution and detection of MTBC in cultures, but they are rapid tools for laboratory TB diagnosi

Objetivos: O estudo objetivou avaliar métodos molecular e imunológico e propor um fluxo de trabalho utilizando-os para a rotina de diagnóstico da tuberculose (TB). Métodos: Foi realizado um estudo transversal retrospectivo, incluindo 121 culturas líquidas de um laboratório de TB localizado no extremo sul do Brasil. Todas as culturas foram positivas para o complexo Mycobacterium tuberculosis (CMTB) por Reação em Cadeia da Polimerase (PCR) in-house para detecção do IS6110, usando DNA extraído pelo método CTAB (PCR-CTAB). Essas culturas foram submetidas a testes mais rápidos que este, o ensaio imunológico MPT64 e a PCR com DNA extraído pelo método de lise térmica (PCR-LT), e estas foram avaliadas para identificação de CMTB usando PCR-CTAB como método de referência. Resultados: A sensibilidade do ensaio MPT64 e da PCR-LT para identificar o CMTB em culturas positivas pela PCRCTAB foi de 73,6% (89/121) e 98,3% (119/121), respectivamente. Propusemos um fluxo de trabalho baseado no uso do ensaio MPT64 em culturas líquidas sugestivas de CMTB e, em caso de resultado negativo, sugerimos a realização de PCR-LT. Sugere-se a PCR-CTAB apenas se os testes mais rápidos forem negativos. Conclusões: Os métodos capazes de confirmar o CMTB em culturas devem continuar sendo padronizados, testados e otimizados para atender aos requisitos ideais de simplicidade, rapidez e eficácia. Os métodos molecular e imunológico avaliados apresentam diferenças na execução e detecção do CMTB em culturas, mas são ferramentas rápidas para o diagnóstico laboratorial da TB.

El dilema en los laboratorios de Koch, ¿Cultivar o no Cultivar? / The Dilemma in Koch's Laboratories, To Cultivate or Not to Cultivate?

La nueva norma técnica para el control y la eliminación de la tuberculosis es un gran avance para el diagnóstico de este microorganismo en Chile. Actualmente la principal técnica microbiológica para el diagnóstico de laboratorio es la biología molecular, que reduce el tiempo del resultado a tan solo un par de horas. La normativa actual indica que en el paciente caso presuntivo de tuberculosis (CPT) la técnica exclusiva a realizar es Biología molecular. La literatura indica que la detección a través de amplificación de material genético de la micobacteria tiene un límite de detección de 15,6 UFC/ ml, por tanto, todas las muestras bajo ese límite umbral potencialmente podrían no ser diagnosticadas bajo esta estructura emanada por el ministerio de Salud en Chile. Nuestra recomendación es continuar con el estudio de cultivo en medios líquidos o sólidos para todas las muestras hasta obtener literatura que avale lo contrario

The new technical standard for the control and elimination of tuberculosis in Chile is a great advance for the diagnosis of this microorganism. Currently the main microbiological technique for laboratory diagnosis is PCR, which reduces the time to result to just a couple of hours. The current regulations indicate that in the patient with a presumptive case of tuberculosis (CPT) t he exclusive technique to be performed is PCR. The literature indicates that the detection through amplification of genetic material of the mycobacterium has a detection limit of 15.6 CFU/ml, therefore, all samples under this threshold limit could potentially not be diagnosed under this structure emanated by the Ministry of Health in Chile. Our recommendation is to continue with the study of culture in liquid or solid media for all samples until literature confirms otherwise

Tuberculosis cutánea esporotricoide en la Amazonía peruana: reporte de caso / Sporotrichoid cutaneous tuberculosis in the peruvian amazon: case report

RESUMEN La tuberculosis cutánea es una presentación rara de la infección por Mycobacterium tuberculosis. Se presenta el caso de una mujer sin antecedentes médicos de importancia, con un tiempo de enfermedad de año y medio, caracterizado por lesiones tipo esporotricoide, con diseminación linfocutánea en miembro superior derecho, de evolución lentamente progresiva. Se realizó un estudio histopatológico encontrándose células gigantes tipo Langhans y escasa necrosis. El paciente recibió terapia de esquema sensible antituberculoso, con evolución favorable.

ABSTRACT Cutaneous tuberculosis is a rare presentation of Mycobacterium tuberculosis infection. We present the case of a woman without important medical history, with a disease period of one year and a half, characterized by sporotrichoid-like lesions, with lymphocutaneous dissemination in the right upper limb, and with slowly progressive evolution. The histopathological tests revealed Langhans type giant cells and scarce necrosis. The patient received therapy with a sensitive antituberculous scheme, and evolved favorably.

Imunologia da tuberculose: uma revisão narrativa da literatura / Tuberculosis immunology: a narrative literature review

A resposta imune desenvolvida pelo hospedeiro contra o Mycobacterium tuberculosis é considerada de natureza complexa e multifacetada. Esta interação bacilo-hospedeiro resulta, na maioria das vezes, em uma infecção latente assintomática, podendo ou não evoluir para a forma ativa da tuberculose (TB). O presente estudo objetivou atualizar e sumarizar o conhecimento científico acerca dos mecanismos imunológicos associados à infecção e sua progressão para a TB ativa. Trata-se de uma revisão narrativa, realizada a partir do levantamento bibliográfico de artigos científicos indexados nas bases de dados PubMed/MEDLINE e SciELO, nos últimos 20 anos. Nas últimas décadas, a caracterização de linfócitos Tγδ, MAIT, iNKT e outra células T CD1 restritas proporcionaram um maior entendimento do papel da imunidade inata na infecção pelo bacilo. A migração de linfócitos T CD4+ produtores de IFN-g, TNF-α e de outras moléculas solúveis, promove o recrutamento e formação do granuloma, estrutura que beneficia tanto o hospedeiro quanto o bacilo. Eventualmente, um desequilíbrio nesta complexa rede de interação, resulta em uma resposta inflamatória exacerbada que contribui para o desenvolvimento de um granuloma necrótico. Por fim, a exaustão da resposta imune local frente à contínua exposição ao bacilo, associada ao perfil anti-inflamatório dos linfócitos Th2 e linfócitos Treg, favorecem a inativação funcional e, consequentemente, o desenvolvimento da doença ativa. A resposta imunológica é crucial para o desenvolvimento da infecção por M. tuberculosis. Portanto, estudos que possibilitem uma maior compreensão sobre a interação bacilohospedeiro podem viabilizar o desenvolvimento de novos métodos diagnósticos, estratégias terapêuticas e, sobretudo, avanços no desenvolvimento de imunobiológicos.

The immune response developed by the host against Mycobacterium tuberculosis is considered a complex and multifaceted nature. This host-bacillus interaction, which in most cases results in an asymptomatic latent infection that may or may not evolve to the development of active pulmonary tuberculosis (TB). The present study aimed to update and summarize the current scientific knowledge regarding the immunological mechanisms associated with infection and the development of active disease. This is a narrative review, based on scientific articles indexed in the PubMed/ MEDLINE and SciELO databases over the last 20 years. In recent decades, the characterization of Tγδ lymphocytes, MAIT, iNKT and CD1-restricted T cells has provided a better understanding of the role of innate immunity in bacilli infection. The migration of T CD4+ lymphocytes that produce IFN-γ, TNF-α and other soluble molecules, promotes the recruitment and formation of the granuloma, a structure that benefits both the host and the bacillus. Eventually, an imbalance in this complex interaction network results in an exacerbated inflammatory response that contributes to the development of a necrotic granuloma. Finally, exhaustion of the local immune response due to continuous exposure to the bacillus, associated with the anti-inflammatory profile of Th2 lymphocytes and Treg lymphocytes, favor functional inactivation and, consequently, the development of active disease. The immune response is crucial for the development of M. tuberculosis infection. Therefore, studies that enable a greater understanding of the hostbacillus interaction may enable the development of new diagnostic methods, therapeutic strategies and, above all, advances in the development of immunobiologicals.

GeneXpert como método de diagnóstico de la tuberculosis en Santiago de Cuba / GeneXpert as diagnostic method of tuberculosis in Santiago de Cuba

Introducción: La tuberculosis es un problema de salud en el mundo, por lo que se necesitan métodos como el GeneXpert para realizar un diagnóstico rápido y seguro. Objetivo: Determinar la precisión del GeneXpert como método para el diagnóstico de la tuberculosis en Santiago de Cuba en relación con los estudios tradicionales. Métodos: Se efectuó un estudio descriptivo y transversal desde diciembre de 2018 hasta igual mes de 2019 de 31 pacientes a quienes se les realizaron los 3 métodos para el diagnóstico de la tuberculosis. Se utilizaron variables, tales como edad, sexo, muestras estudiadas, positividad de los métodos utilizados, así como concordancia entre el GeneXpert y el cultivo mediante el índice de Kappa. Resultados: En la serie predominaron los pacientes mayores de 50 años (48,4 %), el sexo masculino (66,7 %) y el esputo como muestra más representativa (80,6 %); asimismo, el cultivo resultó positivo en 32,3 % y el GeneXpert en 22,6 %. En tanto, se diagnosticó tuberculosis en 11 pacientes y el índice de Kappa fue de 0,58. Conclusiones: La determinación de GeneXpert en el diagnóstico de la tuberculosis es precisa, dada su sensibilidad y especificidad altas en relación con los estudios tradicionales de esputo y cultivo, lo cual se confirmó por la elevada concordancia del índice de Kappa.

Introduction: Tuberculosis is a health problem worldwide, reason why methods as the GeneXpert are needed to carry out a quick and safe diagnosis. Objective: To determine the accuracy of GeneXpert as a method for the diagnosis of tuberculosis in Santiago de Cuba in connection with the traditional studies. Methods: A descriptive and cross-sectional study was carried out from December, 2018 to the same month in 2019 of 31 patients to whom the 3 methods for the diagnosis of tuberculosis were carried out. Some variables were used, such as age, sex, studied samples, positivity of the used methods, as well as consistency between the GeneXpert and the culture by means of the Kappa index. Results: In the series there was a prevalence of the patients over 50 years (48.4 %), male sex (66.7 %) and sputum as more representative sample (80.6 %); also, the culture was positive in 32.3 % and GeneXpert in 22.6 %. As long as, tuberculosis was diagnosed in 11 patients and the Kappa index was 0.58. Conclusions: The determination of GeneXpert in the diagnosis of tuberculosis is necessary, given its high sensibility and specificity in connection with the traditional studies of sputum and culture, which was confirmed due to the high consistency of the Kappa index.

Caracterización clínica y microbiológica de pacientes con trazas en Xpert MTB/RIF Ultra / Clinical and microbiological characterization of patients with Xpert MTB/RIF Ultra traces

INTRODUCCIÓN: El Xpert MTB/RIF Ultra (Ultra) ha mejorado dramáticamente el diagnóstico de la tuberculosis (TBC). Con él ha nacido la categoría de trazas, que es la menor carga bacilar detectable por este examen. OBJETIVO: Describir las características clínicas de los pacientes con presencia de trazas en el Ultra y evaluar la confirmación de la TBC como diagnóstico clínico. MATERIALES Y MÉTODOS: Estudio descriptivo de serie de casos. Se extrajo la información de fichas clínicas de pacientes con positividad a trazas. Se confrontaron datos clínicos, microbiológicos e histopatológicos. RESULTADOS: Se analizaron 21 pacientes. La edad promedio fue de 52 años. Todos los casos presentaron baciloscopias negativas. Cuatro cultivos en medio líquido MGIT fueron positivos, dos en pleura parietal, uno en líquido pleural y otro en expectoración. En pleura parietal, tres casos presentaron granulomas con necrosis caseosa y un granuloma esbozos de necrosis. En tejido pulmonar se observaron dos casos con granulomas con esbozos de necrosis y dos con granulomas no necrotizantes. Tres pacientes tenían el antecedente de TBC previa, se interpretó la positividad de trazas en ellos como falsos positivos. Finalmente se diagnosticaron 13 casos como TBC activa, donde cinco de ellos fueron TBC pleurales. La mayor concordancia clínica, microbiológica e histopatológica fue en muestras de líquido y tejido pleural. DISCUSIÓN: Se debe interpretar con cautela los hallazgos de esta prueba en muestras de vía aérea; el análisis multidisciplinario (clínica, imágenes, microbiología, histología) es crucial en las decisiones de nuestras conductas clínicas futuras. El hallazgo de trazas en pleura tiene, a nuestro parecer, un alto valor diagnóstico en el estudio de la tuberculosis en esta localización.

INTRODUCTION: Xpert MTB/RIF Ultra has dramatically changed the diagnosis of tuberculosis. A new category called traces appeared, which is the smallest amount of bacillar load detectable. OBJECTIVE: Describe the clinical characteristics of patients that present traces in Xpert MTB/RIF Ultra test, and to evaluate the confirmation of tuberculosis as clinical diagnosis. METHODS: We perform a descriptive case series study. Information was recovered from clinical records of patients with positive test for traces. Clinical, histopathological and microbiological results were confronted. RESULTS: Twenty one patients were analyzed. The mean age was 52 years-old. All cases had negative smear microscopy and four MGIT cultures were positive, two in pleural fluid and another in sputum. In parietal pleura, three cases presented granulomas with caseous necrosis, and one showed granuloma with very little necrosis. In pleural tissue we observed two cases of granulomas with traces of necrosis and two with non-necrotizing granulomas. Three patients had history of previous tuberculosis and positive traces, the test was interpreted as a false positive result. Finally, active tuberculosis was diagnosed in 13 cases, and five of them were pleural tuberculosis. The highest clinical, microbiological and histopathological agreement was in fluid and pleural tissue samples. DISCUSSION: The findings of Xpert MTB/RIF Ultra in airway samples must be interpreted carefully. Multi-disciplinary analysis is crucial in future clinical decisions. The finding of traces in pleura has, in our opinion, a high diagnostic value in the study of tuberculosis in this location.

Clinical and epidemiological features of tuberculosis isolated from critically ill patients / Tuberculosis en la unidad de cuidados intensivos: características clínicas y epidemiológicas

Human tuberculosis is still a major world health concern. In Uruguay, contrary to the world trend, an increase in cases has been observed since 2006. Although the incidence of MDR-resistant strains is low and no cases of XDR-TB were registered, an increase in the number of patients with severe tuberculosis requiring critical care admission was observed. As a first aim, we performed the analysis of the genetic structure of strains isolated from patients with severe tuberculosis admitted to an intensive care unit. We compared these results with those corresponding to the general population observing a statistically significant increase in the Haarlem genotypes among ICU patients (53.3% vs 34.7%; p;<;0.05). In addition, we investigated the association of clinical outcomes with the genotype observing a major incidence of hepatic dysfunctions among patients infected with the Haarlem strain (p;<;0.05). The cohort presented is one of the largest studied series of critically ill patients with tuberculosis.

La tuberculosis (TB) aún representa un problema mayor de salud pública. En Uruguay, contrariamente a la tendencia mundial, se ha observado un incremento en el número de casos desde 2006. Aunque la incidencia de casos de multidrogorresistencia (MDR) es baja y no se han reportados casos de resistencia a fármacos de primera y segunda línea de tratamiento (XDR), se ha observado un incremento en el número de casos con TB grave, que requieren internación en unidad de terapia intensiva (CTI). Como primer objetivo del presente trabajo, se analizó la estructura genética de cepas de Mycobacterium tuberculosis aisladas de pacientes internados en CTI. Comparamos estos resultados con los obtenidos con cepas circulantes en la comunidad. Observamos un incremento estadísticamente significativo del genotipo Haarlem en los pacientes internados en CTI (53,3 vs. 34,7%; p;<;0,05). Además, investigamos la asociación del desenlace clínico con el genotipo, y encontramos una mayor incidencia de disfunción hepática en los pacientes infectados con la cepa Haarlem (p;<;0,05). La cohorte presentada en este trabajo corresponde a una de las series con mayor número de pacientes con tuberculosis que requirieron internación en CTI.

Evaluación de la susceptibilidad in vitro a esparteína, en cuatro cepas de Mycobacterium tuberculosis / Evaluation of in vitro susceptibility to sparteine in four strains of Mycobacterium tuberculosis

RESUMEN La esparteína es un alcaloide con actividad bacteriostática sobre el género Mycobacterium. El objetivo de este trabajo fue evaluar la acción antimicrobiana de la esparteína en el crecimiento de cuatro cepas ATCC de Mycobacterium tuberculosis (susceptible, resistente a isoniazida, resistente a rifampicina y multidrogorresistente) in vitro. La evaluación de la actividad bactericida del sulfato de esparteína se realizó a través de una adaptación del método de ensayo de cultivo y susceptibilidad a medicamentos antituberculosos mediante observación microscópica (MODS, por sus siglas en inglés), según el protocolo descrito en el manual técnico elaborado por el Instituto Nacional de Salud. Los resultados demuestran que a concentraciones de 25; 50 y 100 mM de sulfato de esparteína, no se desarrollan unidades formadoras de colonia en las cuatro cepas evaluadas de Mycobacterium tuberculosis. Los resultados demuestran el potencial efecto antimicrobiano in vitro de la esparteína en la tuberculosis multidrogorresistente.

ABSTRACT Sparteine is an alkaloid with bacteriostatic activity on the genus Mycobacterium. The aim of this study was to evaluate the antimicrobial activity of sparteine on the growth of 4 ATCC strains of Mycobacterium tuberculosis (susceptible, resistant to isoniazid, resistant to rifampicin and multidrug-resistant) in vitro. Validation of bactericidal activity of sparteine sulfate was carried out through an adaptation of the Microscopic-Observation Drug-Susceptibility (MODS) method according to the guidelines of the Peruvian National Health Institute. The results demonstrate that at concentrations of 25; 50 and 100 Mm of sparteine sulfate, there is no development of colony-forming units in any of the 4 evaluated strains. Our results demonstrate the potential in vitro antimicrobial effect of sparteine on multidrug-resistant tuberculosis.

Eritema indurado de Bazin como una reactivación de una tuberculosis latente en una paciente con tratamiento biológico anti-factor de necrosis tumoral / Erythema induratum of Bazin as a reactivation of latent tuberculosis infection in a patient receiving anti-tumor necrosis factor treatment

Resumen El eritema indurado de Bazin es una tuberculosis cutánea rara, considerada una tuberculide o reacción de hipersensibilidad a Mycobacterium tuberculosis. El tratamiento con agentes biológicos es un factor de riesgo conocido para la reactivación de tuberculosis, especialmente en áreas de alta incidencia como Latinoamérica, por lo que existen protocolos de búsqueda y tratamiento antes del inicio de este tipo de terapias. Se presenta un caso clínico de eritema indurado de Bazin como reactivación de una infección tuberculosa latente en una paciente con artritis reumatoide que recibía tratamiento con golimumab.

Abstract Erythema induratum of Bazin is a rare form of cutaneous tuberculosis, considered as part of the spectrum of tuberculids or hipersensitivity reactions to Mycobacterium tuberculosis. Treatment with biologic agents is a known risk factor for tuberculosis reactivation, especially in areas of high incidence like Latin America, which is why screening and treatment protocols must be followed before these therapies are initiated. We present a case of erythema induratum of Bazin as a reactivation of latent tuberculosis infection in a patient with rheumatoid arthritis treated with golimumab.

Heparin-binding hemagglutinin as a composition antigen of tuberculosis vaccine exerts protective immune effects by inducing IL-17 / 中华预防医学杂志

To explore the protective immune effect induced by mucosal delivery heparin-binding hemagglutinin (HBHA)-a candidate vaccine antigen of Mycobacterium tuberculosis. Female C57BL/6 mice were between 6 and 8 weeks of age before experimental use. Thirty mice received different immunization strategies and were randomly divided into the control group, the early secreting antigen target-6 (ESAT-6) intranasal immunization group, the HBHA intranasal immunization group, the BCG priming PBS control group, or BCG priming HBHA boost group, 6 mice in each group. In order to analyzed the immune effect, the concentrations of plasma Interleukin-17A (IL-17A) and other cytokines were measured by ELISA. Quantitative real-time PCR analyses were performed to detect the relative quantity (RQ) mRNA of IL-17A in the lung. The lung tissue sections were stained to detect the formation of the tertiary lymphoid structures. The chemokines contributed to formation of the tertiary lymphoid structures were also measured. Flow cytometry was used to detect the frequency of Th1 and Th17 cells in the system. Sixty mice in the BCG priming PBS control group and the BCG priming HBHA boost group were sacrificed at different time points after infection to count the lung bacterial burden. The concentrations of plasma IL-17A and relative quantity of lung IL-17A mRNA were highest in the BCG priming HBHA boost group [(14.76±4.73) pg/mL,RQ (12.27±6.71)], which was significantly higher than the control group [(5.57±2.95) pg/mL,RQ (1.30±0.97)] (t=4.213, P<0.001; t=5.984, P<0.001), and also significantly higher than the BCG priming PBS control group [(6.81±2.18) pg/mL,RQ (1.44±1.16)] (t=3.646 P=0.001; t=6.185 P<0.001). Compared with the BCG priming PBS control group (0.38±0.38)% the frequency of spleen Th17 cells were also significantly increased (t=-0.280 , P=0.048) in the BCG-primary HBHA boost group (1.02±0.34)%. In addition, HBHA boosting could promote better formation of the tertiary lymphoid structures in the lung, and decrease the bacterial load on the early stage after BCG challenge. Collectively, mucosal delivery of HBHA can effectively enhance the protective effect after BCG vaccination, and it is a potential candidate vaccine component.

Detection of drug resistance genes of mycobacterium tuberculosis by rolling circle amplification technique with multicolor fluorescent probes / 中华预防医学杂志

The aim of this study was to construct a simple, rapid and ultra-sensitive optical biosensing technique based on rolling circle amplification (RCA), and to apply it to multiple detection of drug-resistant genes of mycobacterium tuberculosis. The common mutation sites of isoniazid, rifampicin and streptomycin resistance genes are katG315 (AGC➝ACC), rpoB531 (CAC➝TAC) and rpsL43 (AAG➝AGG). For these three gene sites, from February 2020 to May 2021, in the Department of Laboratory Medicine of the First Affiliated Hospital of Army Military Medical University, the padlock probe (PLP), primers and capture probes were designed. And a solid-phase RCA constant temperature amplification reaction system based on magnetic beads was constructed and the experimental parameters were optimized. The RCA products were accurately captured by the multicolor fluorescent probes (Cy3/Cy5/ROX), and the single-tube multiple detection of three mutation genes was realized. The sensitivity, specificity and linear range of this method were further verified. The results showed that the response range of katG315 in the same reaction system ranged from 1.0 pmol/L to 0.1 nmol/L. The response range of rpoB531 and rpsL43 ranged from 1.0 pmol/L to 50.0 pmol/L and 1.0 pmol/L to 20.0 pmol/L, and the method had good specificity and sensitivity, and could accurately identify single base mutations in mixed targets, with the minimum detection limit as low as 1.0 pmol/L. The recoveries of simulated serum samples were 95.0%-105.2%. In conclusion, the constant temperature amplification multiple detection method constructed in this study can quickly realize the single-tube multiple detection of three drug resistance mutation sites. This technology is low-cost, simple and rapid, and does not rely on large equipment, providing a new analysis method for pathogen drug resistance gene detection.

Case report : Calvarial tuberculosis

@#<p>Skeletal tuberculosis accounts for 1-3% of TB cases, and of these only 0.2-1.3% had calvarial involvement. 1 Calvarial TB is most likely secondary to a primary focus.  Diagnosis is confirmed through findings of Mycobacterium tuberculosis via microbiological, histopathological or cytopathological methods. This case report presents<br />Primary Calvarial Tuberculosis in a five-year old male presenting with multiple cranial masses and initial diagnosis of Langerhans cell histiocytosis (LCH).</p>

Clinical neglect of aspergillosis in pulmonary tuberculosis co-infection: a case report of avoidable mortality in a resource-constrained setting

Background: Pulmonary aspergillosis (PA) is common among patients with tuberculosis (TB). With both infections presenting with similar clinical and radiologic features, diagnosis of PA is often made too late or missed completely due to lack of clinical suspicion and poor diagnostic laboratory capacity for mycotic infections prevalent in our settings. We present a case of preventable mortality caused by delayed diagnosis and treatment of PA in a patient with pulmonary TB (PTB). Case presentation: A 13-year-old female was diagnosed and treated for PTB, having received anti-TB regimen for 8 months in a mission hospital from where she was referred due to worsening cough, chest pain and progressive breathlessness. The patient was re-assessed and investigated, with GeneXpert detecting Mycobacterium tuberculosis, susceptible to rifampicin. Diagnosis of pulmonary tuberculosis complicated by right pneumothorax was made indicating an emergency thoracotomy and chest tube insertion and continuation of the first line anti-TB regimen. At about 2 weeks into admission, patients had features of superimposed acute bacterial sepsis with fever becoming high grade, marked neutrophilia with toxic granulation and elevated sepsis biomarker, and this necessitated empiric antibiotic treatment with parenteral meropenem and vancomycin. However, the patient only had mild clinical improvement following which there was progressively worsening respiratory symptoms and massive haemoptysis. Result of sputum fungal study was available on admission day 20 and revealed a growth of Aspergillus flavus. Treatment with intravenous voriconazole was however commenced rather late when the fungal respiratory disease could no longer be remedied. The patient died on admission day 23. Conclusion: Diagnosis of PA in patients with background TB is often made too late to guarantee timely and effective antifungal treatment with negative consequences on patients' outcomes. Improving clinical and laboratory capacities is essential to reducing mortality from PA in healthcare facilities.

Sobrevida en una cohorte con diagnóstico de tuberculosis en Colombia / Survival at cohort diagnosed with tuberculosis in Colombia

Introducción: La tuberculosis es una patología infecciosa crónica cuya incidencia es elevada en países en vía de desarrollo, sin embargo, es limitada la información y los estudios que analizan la mortalidad y sobrevida a largo plazo. Metodología: estudio de cohorte retrospectivo, en pacientes con diagnóstico de tuberculosis mayores de 18 años, el ingreso fue de manera consecutiva hasta completar el periodo de estudio. Se analizó la sobrevida y mortalidad a través del estimador Kaplan ­ Meier por la prueba de log Rank. Resultados: ingresaron 329 sujetos, la mortalidad a los 30 días fue de 11,9% y al año del 24,6%, la tuberculosis pulmonar fue el tipo más frecuente con en el 70,2%. Los hallazgos al examen físico relacionados con mortalidad fueron la caquexia (p<0,001) y el edema en extremidades (p<0,001). La sobrevida general fue del 87,2% a los 30 días y del 72,9% al año. En los pacientes con tuberculosis pulmonar la sobrevida fue del 85,8% a los 30 días y del 72,8% al año. Conclusión: La tasa de sobrevida a un año en pacientes hospitalizados por tuberculosis es baja, la edad avanzada, desnutrición, PaO2/FiO2 menor de 300, proteína c reactiva mayor de 45 mg/dL, enfermedad cerebrovascular y enfermedad vascular periférica fueron variables que se asociaron con una mayor mortalidad(AU)

Background: Tuberculosis is a chronic infectious pathology whose incidence is high in developing countries, however, information and studies that analyze mortality and long-term survival are limited. Methodology: retrospective cohort study, in patients with a diagnosis of tuberculosis older than 18 years, admission was consecutive until completing the study period. Survival and mortality were analyzed using the Kaplan-Meier estimator by the log Rank test. Results: 329 subjects were admitted, mortality at 30 days was 11.9% and at one year 24.6%, pulmonary tuberculosis was the most frequent type with 70.2%. Physical examination findings related to mortality were cachexia (p<0.001) and extremity edema (p<0.001). Overall survival was 87.2% at 30 days and 72.9% at one year. In patients with pulmonary tuberculosis, survival was 85.8% at 30 days and 72.8% at one year. Conclusion: The one-year survival rate in patients hospitalized for tuberculosis is low, advanced age, malnutrition, PaO2/FiO2 less than 300, c-reactive protein greater than 45 mg/dL, cerebrovascular disease and peripheral vascular disease were variables that were associated with higher mortality(AU)

Prospecção de novos candidatos a fármacos antituberculose: uma abordagem in vitro / Prospecting new candidates for antituberculosis drugs: an in vitro approach

A tuberculose (TB) é uma doença infecciosa causada pelo Mycobacterium tuberculosis que apresenta alta morbidade e mortalidade. Atualmente, a tuberculose afeta 9,9 milhões de pessoas em todo o mundo e é responsável por cerca de 1,3 milhões de mortes, sendo um sério problema de Saúde Pública global. O tratamento da TB é composto por uma associação de fármacos e dura seis meses. A frequência de doentes infectados com isolados resistentes aos fármacos de primeira linha, principalmente os isolados multirresistentes, é uma ameaça mundial que caracteriza um importante problema de saúde pública no controle da TB em vários países. Existem poucos medicamentos para o tratamento da TB muti-resistente e com base nesta problemática o presente estudo visou a avaliação do potencial farmacológico in vitro de fármacos aprovados, utilizando a abordagem de reposicionamento de fármacos. A partir de uma triagem in vitro de 89 fármacos, foram selecionados os que apresentaram atividade antituberculose para a avaliação da concentração inibitória mínima, os fármacos foram avaliados em exposição a 23 isolados do complexo Mycobacterium tuberculosis e a cepa ATCC H37 RV 27294 in vitro utilizando o ensaio de resazurina. Além disso, foram realizados ensaios de citotoxicidade contra células de mamíferos NCTC L929 em cultura, determinando-se os Índices de Seletividade in vitro. Dentre os 89 fármacos testados, quatro (amiodarona, flunarizina, ivermectina e pentamidina) apresentaram atividade antituberculose a 10 µM. A Concentração Inibitória Mínima (CIM) 90% da amiodarona resultou em 5-10 µM, flunarizina 10 µM, ivermectina 0,15 ­ 10 µM e pentamidina 1,25-10 µM. Os índices de seletividade observados para a amiodarona foram de 4,21 ­ 8,43, flunarizina >20, ivermectina 1,17 ­ 78,47 e pentamidina 14,40 ­ 115,2. Com base na atividade e citotoxicidade da amiodarona, foi realizado um ensaio fluorimétrico utilizando Sytox Green para avaliar a permeabilidade de membrana plasmática do complexo M. tuberculosis na presença deste fármaco. Foi observado que a amiodarona não alterou a permeabilidade da membrana plasmática do Complexo M. tuberculosis, tendo seu mecanismo de ação letal por outra via.

GenoType MTBDRsl for detection of second-line drugs and ethambutol resistance in multidrug-resistant Mycobacterium tuberculosis isolates at a high-throughput laboratory

We assessed the performance of MTBDRsl for detection of resistance to fluoroquinolones, aminoglycosides/cyclic peptides, and ethambutol compared to BACTEC MGIT 960 by subjecting simultaneously to both tests 385 phenotypically multidrug-resistant-Mycobacterium tuberculosis isolates from Sao Paulo, Brazil. Discordances were resolved by Sanger sequencing. MTBDRsl correctly detected 99.7% of the multidrug-resistant isolates, 87.8% of the pre-XDR, and 73.9% of the XDR. The assay showed sensitivity of 86.4%, 100%, 85.2% and 76.4% for fluoroquinolones, amikacin/kanamycin, capreomycin and ethambutol, respectively. Specificity was 100% for fluoroquinolones and aminoglycosides/cyclic peptides, and 93.6% for ethambutol. Most fluoroquinolone-discordances were due to mutations in genome regions not targeted by the MTBDRsl v. 1.0: gyrA_H70R and gyrB_R446C, D461N, D449V, and N488D. Capreomycin-resistant isolates with wild-type rrs results on MTBDRsl presented tlyA mutations. MTBDRsl presented good performance for detecting resistance to second-line drugs and ethambutol in clinical isolates. In our setting, multidrug-resistant. isolates presented mutations not targeted by the molecular assay.