GenoType MTBDR plus 1.0 ® para la detección de resistencia cruzada entre isoniacida y etionamida en aislamientos de Mycobacterium tuberculosis multirresistentes / GenoType MTBDR plus 1.0 ® for the detection of cross-resistance between isoniazide and ethionamide in isolates of multidrug-resistant Mycobacterium tuberculosis

Introducción. Una parte de los aislamientos de Mycobacterium tuberculosis multirresistente también presenta resistencia a la etionamida. Es importante determinar si la resistencia a la isoniacida es independiente o se cruza con la resistencia a la etionamida, ya que si sucede lo segundo habría que reevaluar el tratamiento antituberculoso de segunda línea. La prueba molecular GenoType MTBDR plus ® detecta las mutaciones asociadas con la resistencia a isoniacida y podría detectar la resistencia cruzada a la etionamida. Objetivo. Evaluar la prueba GenoType MTBDR plus ® y comparar su desempeño con el de la secuenciación, en la detección de mutaciones en el gen katG y en el promotor inhA en aislamientos clínicos de M. tuberculosis multirresistente. Materiales y métodos. Se utilizaron el estuche comercial GenoType MTBDR plus 1.0 ® y la secuenciación para evaluar mutaciones en el gen katG y en el promotor inhA en 30 aislamientos de M. tuberculosis multirresistente con resistencia a la etionamida. La cepa de laboratorio H37Rv y tres aislamientos sensibles a los medicamentos de primera línea, sirvieron de control. Resultados. Al comparar los resultados de la secuenciación y de la prueba GenoType MTBDR plus ® , el índice kappa fue de 1. Todos los aislamientos resistentes a la isoniacida y la etionamida tenían las mutaciones detectadas con la prueba GenoTypeMTBDR plus ® en el gen katG, y 40 % de ellos, las detectadas en el promotor inhA. Mediante secuenciación se encontraron, además, mutaciones en katG en posiciones diferentes a las detectadas por la prueba GenoType MTBDR plus ® . Conclusión. La prueba GenoTypeMTBDR plus ® tiene la capacidad de detectar rápidamente la resistencia a isoniacida. Además, los resultados del estudio sugieren que también podría utilizarse como prueba de tamización para detectar la resistencia cruzada a etionamida.

Introduction: A variable proportion of isolates of multidrug-resistant Mycobacterium tuberculosis also presents resistance to ethionamide. It is important to determine whether resistance to isoniazid is independent or crossed with resistance to ethionamide, given that this could lead to the re-evaluation of second-line anti-tuberculosis treatment. The GenoType MTBDR plus ® molecular test is used for the detection of MDR-MTB, as it identifies mutations associated with resistance to isoniazide and could detect cross-resistance with ethionamide. Objective: To evaluate the performance of GenoType MTBDR plus ® in comparison with sequencing in the detection of mutations in gene katG and promotor inhA in clinical isolates of multidrug-resistant M. tuberculosis . Materials and methods: The GenoType MTBDR plus 1.0 ® commercial kit and sequencing were used to evaluate mutations in gene katG and promotor inhA in 30 multidrug-resistant M. tuberculosis isolates that were resistant to ethionamide. The laboratory strain H37Rv and three pan-sensitive isolates acted as controls. Results: The kappa index for the comparison between the results of sequencing and GenoType MTBDR plus ® was 1. All the isolates resistant to isoniazid and ethionamide had the mutations detected by GenoTypeMTBDR plus ® in the katG gene and 40% of them in promotor inhA. Sequencing also revealed katG mutations in positions different to those detected by GenoType MTBDR plus ® . Conclusion: GenoType MTBDR plus ® is able to detect resistance to isoniazid rapidly. Our results suggest that it could also be used to screen for cross-resistance with ethionamide.

Comparison between serum and urinary sulphadimidine acetylation as predictors of isoniazid acetylator status in patients with pulmonary tuberculosis.

The acetylator status of 200 pulmonary tuberculosis patients was determined by sulphadimidine (SDM) acetylation in (1) urine and (2) serum. In both estimations, distribution of the patients with regard to the acetylation rates was found to be bimodal. Based on the percent frequency distribution of acetylated SDM, the antimode for urine and serum was 65% for the former and 45% for the latter. Patients acetylating above 65% or 45% of administered SDM were taken as rapid acetylators, while those acetylating less, were slow acetylators. The ratio of slow to rapid acetylators in urine and serum was 58 : 42 and 66 : 34 respectively. The renal clearance of acetylated fraction of SDM was considerably greater as compared to that of the unacetylated fraction. Agreement between the two methods as evaluated by the kappa statistic was 0.71. The determination of the acetylator status by the SDM acetylation test in the urine is simple to perform and has acceptable accuracy. It may be used as an acceptable substitute to serum for estimating the isoniazid inactivation status which is of considerable importance when patients are treated with daily/ intermittent/once-weekly drug regimens for tuberculosis.

Isoniazid acetylating phenotype in patients with paracoccidioidomycosis and its relationship with serum sulfadoxin levels, glucose-6-phosphate dehydrogenase and glutathione reductase activities

Os autores avaliaram o fenótipo acetilador da isoniazida, hematócrito, hemoglobina, atividade da glicose-6-fosfato desidrogenase, glutationa redutase e os níveis séricos de sulfadoxina de 39 doentes com paracoccidioidomicose, sendo 33 do sexo masculino e 6 do feminino, com idades compreendidas entre 17 e 58 anos. Vinte e um (53,84 por cento) doentes apresentaram fenótipo acetilador lento e 18(46,16 por cento) rápido. A atividade da glicose-6-fosfato desidrogenase (G6PD) esteve diminuída em 5(23,80 por cento) acetiladores lentos e 4(22,22 por cento) rápidos. A atividade da glutationa redutase esteve diminuida em 14 (66,66 por cento) acetiladores lentos e 12 (66,66 por cento) rápidos. Os níveis séricos de sulfadoxina livre e total foram maiores nos acetiladores lentos (p < 0,02). A análise dos resultados permite concluir que os níveis séricos de sulfadoxina relaciona-se com o fenótipo acetilador. Além disso, os níveis estiveram sempre acima de 50µg/ml, níveis estes considerados terapêuticos. Por outro lado, a deficiência de glutationa redutase pode estar relacionada com a má absorçäo intestinal de nutrientes, entre eles riboflavina, vitamina precursora de FAD

Estudio preliminar sobre el fenotipo de acetilador de los amerindios Teribes / Preliminar study on the phenotype of acetylator of Teribe Amerindians

El estudio de la capacidad de acetilación en 62 voluntarios teribes, procedentes de Sieykin, Sieyic y Santa Rosa, demonstró que 48.4% eran acetiladores rápidos y 51.6%, acetiladores lentos. Al comprobar estos resultados con los que obtuvimos en una población cuna, en la cual encontramos que el 78% era de acetiladores rápidos, se observa que tanto la distribución de los sujetos estudiados, según el porcentaje de isoniacida que acetilaron, como el porcentaje de acetiladores lentos y rápidos indican que, en ambos grupos, existen diferencias en la actividad de la N-acetiltransferasa. Estos resultados sugieren que pueden existir diferencias fundamentales entre los diferentes grupos amerindios, en la habilidade de biotransformar los medicamentos

A acetilaçäo da isoniazida na cirrose hepática / Isoniazid acetylation in hepatic cirrhosis

Foi investigado o fenótipo acetilador de isoniazida em 23 pacientes caucasóides com cirrose hepática, por intermédio da avaliaçäo do percentual de acetilisoniazida na urina. A proporçäo de acetiladores lentos entre cirróticos foi bem menor que aquela verificada em um grupo controle de caucasóides. Ao se separar os pacientes quanto ao grau de insuficiência hepática, os dados mostraram que a acetilaçäo da isoniazida é tanto mais lenta quanto maior é a insuficiência hepática. Concluiu-se que a cirrose afeta a capacidade hepática de acetilaçäo da isoniazida, alterando o fenótipo acetilador desse medicamento

A acetilaçäo da isoniazida na síndrome de Gilbert / Isoniazid acetylation in Gilbert's syndrome

O fenótipo acetilador da isoniazida foi investigada em 19 pacientes caucasóides portadores de síndrome de Gilbert, por intermédio da avaliaçäo do percentual de acetil-isoniazida na urina. A proporçäo de acetiladores lentos entre os pacientes com síndrome de Gilbert foi semelhante àquela verificada em um grupo controle de caucasóides. Conclui-se que na síndrome de Gilbert näo há interferência na capacidade hepática de acetilaçäo da isoniazida

Serum concentration and urinary excretion of ethambutol administered alone and in combination with isoniazid in patients of pulmonary tuberculosis.

Ethambutol (20 mg/kg) was administered orally to 10 patients of pulmonary tuberculosis for seven consecutive days at 8 a.m. after over night fast. On 7th day serum levels were measured at 2, 4, 6, 8 and 24 hr intervals and urinary excretion was estimated at 2, 4, 6 and 24 hr following ethambutol administration. Simultaneous administration of isoniazid (300 mg, orally) for next seven days to the same patients significantly raised the serum levels of ethambutol at 4, 6 and 8 hr and the cumulative per cent dose excreted was decreased significantly at 4, 6 and 24 hr. The serum levels and urinary elimination was not significantly different at 2 hr.

Levels of acetyl hydrazines and rate of acetylation of isoniazid in adult tuberculosis patients.

Patients suffering from pulmonary tuberculosis were investigated for the levels of isoniazid (INH) and its metabolites viz. acetyl-INH, mono and diacetyl hydrazines and ammonia. It was observed that 50% of the patients are slow inactivators of INH and almost all show an increase in circulating levels of NH3 at 6 hrs. Mono and diacetyl hydrazine levels in blood and urine were detectable in all the patients up+o 24 hrs though the maximum levels were observed at different intervals after the intake of INH.