RESUMO
INTRODUCTION: The diagnosis of intestinal tuberculosis is challenging even nowadays. This study aims to report the positivity rates of new diagnostic methods such as immunohistochemistry and Real-Time Polymerase Chain Reaction in patients with intestinal tuberculosis, as well as describe the pathological and endoscopic features of intestinal tuberculosis in our population. METHODS: This was a retrospective observational study conducted in patients diagnosed with intestinal tuberculosis, between 2010 to 2023 from the Hospital Nacional Daniel Alcides Carrion and a Private Pathology Center, both located in Peru. Clinical data was obtained, histologic features were independently re-evaluated by three pathologists; and immunohistochemistry and real-time Polymerase Chain Reaction evaluation were performed. The 33 patients with intestinal tuberculosis who fulfilled the inclusion criteria were recruited. RESULTS: Immunohistochemistry was positive in 90.9% of cases, while real-time Polymerase Chain Reaction was positive in 38.7%. The ileocecal region was the most affected area (33.3%), and the most frequent endoscopic appearance was an ulcer (63.6%). Most of the granulomas were composed solely of epithelioid histiocytes (75.8%). Crypt architectural disarray was the second most frequent histologic finding (78.8%) after granulomas, but most of them were mild. CONCLUSION: Since immunohistochemistry does not require an intact cell wall, it demonstrates higher sensitivity compared to Ziehl-Neelsen staining. Therefore, it could be helpful for the diagnosis of paucibacillary tuberculosis.
Assuntos
Imuno-Histoquímica , Reação em Cadeia da Polimerase em Tempo Real , Tuberculose Gastrointestinal , Humanos , Tuberculose Gastrointestinal/diagnóstico , Tuberculose Gastrointestinal/microbiologia , Peru , Masculino , Feminino , Estudos Retrospectivos , Adulto , Pessoa de Meia-Idade , Idoso , Adulto Jovem , Granuloma/diagnóstico , Granuloma/microbiologia , Granuloma/patologia , Mycobacterium tuberculosis/isolamento & purificação , Mycobacterium tuberculosis/genética , Adolescente , Sensibilidade e EspecificidadeRESUMO
Resumen Diversos estudios han evidenciado una resistencia cruzada entre isoniacida y etionamida, 2 de los fármacos utilizados en el tratamiento de la tuberculosis multirresistente.El objetivo del presente estudio fue determinar la resistencia cruzada entre ambos fármacos en aislados de Mycobacterium tuberculosis obtenidos en un hospital de Lima (Perú), conalta proporción de pacientes con tuberculosis. Se calculó la frecuencia de mutaciones asociadas con la resistencia a la isoniacida (INH) evaluando el gen katG y la región promotorainhA mediante la prueba molecular Genotype MTBDRplus v2.0. El método gold standard conocido como agar proporciones en placa (APP) permitió la identificación de resistencia a INH yetionamida. De 107 aislamientos resistentes a INH, 54 fueron multirresistentes (identificadosmediante la prueba Genotype MTBDRplus) y 49 (es decir, el 45,8% del total) también fueronresistentes a etionamida por el método APP. En los aislamientos resistentes a INH, se encontraron mutaciones en el gen katG en el 50,5% (54/107); en la región promotora inhA en el23,3% (25/107), y un 14,0% (15/107) presentaron mutaciones en ambos. Un 12,1% (13/107)fueron resistentes a INH por ausencia de banda wild type y banda de mutación. La mutaciónC-15T en la región promotora inhA presentó una fuerte asociación con la resistencia a etionamida y alcanzó el 73,4% (36/49) de los aislamientos resistentes a dicho fármaco. Los resultadosdel presente estudio sugieren que la identificación de mutaciones relacionadas con resistenciaa INH, sobre todo en la región promotora inhA, podría ser de gran utilidad para identificarla resistencia cruzada a etionamida y mejorar el tratamiento de las personas afectadas portuberculosis.© 2019 Asociacion Argentina de Microbiolog´ía. Publicado por Elsevier Espana, S.L.U. Este es unart´ículo Open Access bajo la licencia CC BY-NC-ND (https://creativecommons.org/licenses/by-nc-nd/4.0/).
Abstract Several studies have shown cross-resistance between isoniazid and ethionamide, 2of the drugs used in the treatment of multidrug-resistant tuberculosis. The objective of this study was to determine the cross-resistance between both drugs in Mycobacterium tuberculosis isolates from a hospital with high incidence of tuberculosis in Lima, Peru. The frequency of mutations to isoniazid in the katG gene and the inhA promoter region was identified by the Genotype MTBDRplus v2.0 molecular test. The gold standard Agar Proportion method (APM) allowed todetect resistance to isoniazid and ethionamide. Of 107 isoniazid-resistant isolates (54 multidrug-resistant isolates identified by the Genotype MTBDRplus test, 45.8% (49/107) were also resistant to ethionamide by the APM. Mutations were found in the katG gene in 50.5% (54/107), in the promoter region inhA in 23.3% (25/107) and 14.0% (15/107) that share both mutations in the resistant isolates to INH. The absence of the wild type and mutation bands indicated that 12.1% (13/107) of the isolates were resistant to INH. The mutation C-15T in the inhA promoter region showed a strong association with resistance to ethionamide in 73.4% (36/49) of the isolates analyzed. The results of the present study suggest that the identification of mutations related to resistance to isoniazid, especially in the inhA promoter region, could be very useful to identify cross-resistance to ethionamide and improve the treatment of individuals suffering from this disease.
Assuntos
Humanos , Tuberculose Resistente a Múltiplos Medicamentos/genética , Etionamida/farmacologia , Isoniazida/farmacologia , Mutação , Mycobacterium tuberculosis/efeitos dos fármacos , Mycobacterium tuberculosis/genética , Antituberculosos/farmacologia , Peru , Interações Medicamentosas , Genótipo , Mycobacterium tuberculosis/isolamento & purificaçãoRESUMO
Several studies have shown cross-resistance between isoniazid and ethionamide, 2of the drugs used in the treatment of multidrug-resistant tuberculosis. The objective of this study was to determine the cross-resistance between both drugs in Mycobacterium tuberculosis isolates from a hospital with high incidence of tuberculosis in Lima, Peru. The frequency of mutations to isoniazid in the katG gene and the inhA promoter region was identified by the Genotype MTBDRplus v2.0 molecular test. The gold standard Agar Proportion method (APM) allowed todetect resistance to isoniazid and ethionamide. Of 107 isoniazid-resistant isolates (54 multidrug-resistant isolates identified by the Genotype MTBDRplus test, 45.8% (49/107) were also resistant to ethionamide by the APM. Mutations were found in the katG gene in 50.5% (54/107), in the promoter region inhA in 23.3% (25/107) and 14.0% (15/107) that share both mutations in the resistant isolates to INH. The absence of the wild type and mutation bands indicated that 12.1% (13/107) of the isolates were resistant to INH. The mutation C-15T in the inhA promoter region showed a strong association with resistance to ethionamide in 73.4% (36/49) of the isolates analyzed. The results of the present study suggest that the identification of mutations related to resistance to isoniazid, especially in the inhA promoter region, could be very useful to identify cross-resistance to ethionamide and improve the treatment of individuals suffering from this disease.
Assuntos
Antituberculosos/farmacologia , Etionamida/farmacologia , Isoniazida/farmacologia , Mutação , Mycobacterium tuberculosis/efeitos dos fármacos , Mycobacterium tuberculosis/genética , Tuberculose Resistente a Múltiplos Medicamentos/genética , Interações Medicamentosas , Genótipo , Humanos , Mycobacterium tuberculosis/isolamento & purificação , PeruRESUMO
Introducción. El aumento de las tasas de resistencia en bacterias uropatógenas y su propagación se han convertido en un problema de salud pública a nivel mundial, cobrando mayor importancia la diseminación de aislamientos resistentes productores de ß-lactamasas de espectro extendido, resultando útil la caracterización molecular de plásmidos y otros elementos genéticos de las bacterias para establecer la relación genética que existe entre ellas y tener datos relevantes que puedan sugerir la dirección de propagación de este tipo de resistencia. El objetivo de nuestra investigación fue determinar los perfiles plasmídicos en aislamientos de Escherichia coli y Klebsiella pneumoniae productoras de ß-lactamasas de espectro extendido de urocultivos del Instituto Nacional de Salud del Niño, Lima, Perú. Métodos. Estudio descriptivo, observacional de corte transversal. Resultados. Se determinaron diferentes patrones electroforéticos, obteniéndose de 1 hasta 7 bandas por muestra y en total 15 bandas de distinto tamaño, variando desde 1.5 kb hasta más de20 kb. Teniendo en cuenta los patrones de huella obtenidos, los aislamientos estudiados se distribuyeron en diferentes grupos o ramas, usando el programa NTSYSpc 2.1 y el algoritmo de agrupamiento UPGMA. Conclusión. No se encontró similitud genética marcada entre los aislamientos de origen comunitario y de origen asociado a atención sanitaria, por lo que no se puede establecer una relación significativa y determinar un origen común entre los mismos.
Introduction. The increase in resistance rates in uropathogenic bacteria and their spread has become in a significant worldwide public health, becoming more important the spread of resistant isolates producing of extended spectrum ß-lactamases, which are useful the molecular characterization of plasmids and other genetic elements of the bacteria to establish the genetic relationship between them, to have relevant data that we can suggest the direction of propagation of this type of resistance. The object of research was to determine the plasmid profiles in isolates of Escherichia coli and Klebsiella pneumoniae producing of extended spectrum ß-lactamases from urine cultures of the National Institute of Child Health, Lima, Peru. Methods. Descriptive, observational cross-sectional study. Results. Were detected different electrophoretic patternsin the samples, obtained from 1-7 bands per sample and a total of 15 bands of different sizes, which varied from 1.5 kb to more than 20 kb. Considering the fingerprint patterns, the isolates analyzed were distributed into different clusters or branches, using the NTSYSpc 2.1 program and the UPGMA clustering algorithm. Conclusions. No genetic similarity was found between isolates of community origin and of origin associated with health care, so it is not possible to establish a meaningful relationship and to determine a common origin among them.