Identificación de Mycobacterium bovis en pacientes con diagnóstico de tuberculosis pulmonar y extrapulmonar.

INTRODUCTION: In Mexico, there is an alarming increase in the number of cases of Mycobacterium bovis infection on pulmonary and extrapulmonary presentations. The lack of timely identification triggers complications and increases mortality. OBJECTIVE: To know the frequency of M. bovis infections in clinical samples of patients with tuberculosis in the mycobacteria laboratory of a reference hospital in Mexico City. METHOD: Prospective, descriptive study. Strains isolated from biological material were studied in Löwestein-Jensen and MGITI960 cultures. M. bovis was identified by amplifying the RD9 fragment with end-point polymerase chain reaction (PCR). RESULTS: Eight-hundred and fifty tuberculosis-diagnosed patients were included; in 441 cases, Mycobacterium tuberculosis was confirmed by positive culture (250 pulmonary, 65 ganglionic, 39 renal, 34 meningeal, 25 miliary, 14 pleural, 8 peritoneal, 4 bone and 2 pericardial cases). Forty-eight strains (10.8%) were typified as M. bovis by amplification of the RD9 fragment with end-point PCR. CONCLUSIONS: M. bovis is not currently thought of a causative agent of tuberculosis, which could be the cause of pharmacological treatment failure. In this study, the main extrapulmonary form was observed to be cervical lymphadenopathy.

INTRODUCCIÓN: En México existe un incremento alarmante de casos de infección pulmonar y extrapulmonar por Mycobacterium bovis. La falta de identificación oportuna deriva en complicaciones y eleva la mortalidad. OBJETIVO: Conocer la frecuencia de infecciones por Mycobacterium bovis en muestras clínicas de pacientes con tuberculosis, identificadas en el laboratorio de micobacterias en un hospital de concentración de la Ciudad de México. MÉTODO: Estudio prospectivo, descriptivo. Se estudiaron cepas aisladas de material biológico en cultivos Löwestein-Jensen y MGITI960. La identificación de Mycobacterium bovis se realizó mediante la amplificación del fragmento RD9 por PCR punto final. RESULTADOS: Se incluyeron 850 pacientes con diagnóstico de tuberculosis, en 441 casos se confirmó Mycobacterium tuberculosis por cultivo positivo (250 casos pulmonares, 65 ganglionares, 39 renales, 34 meníngeos, 25 miliares, 14 pleurales, ocho peritoneales, cuatro óseos y dos pericárdicos). Se tipificaron 48 cepas (10.8 %) como Mycobacterium bovis por amplificación del fragmento RD9 por PCR punto final. CONCLUSIONES: Actualmente no se piensa en Mycobacterium bovis como agente causal de tuberculosis, lo que pudiera ser la causa del fracaso del tratamiento farmacológico. En este estudio se observó que la principal forma extrapulmonar es la linfadenopatía cervical.

Nontuberculous mycobacteria in cervical lymphadenopathies of HIV positive and HIV-negative adults / Micobacterias no tuberculosas en linfadenopatías cervicales de adultos VIH positivos y VIH negativos

Background: Tuberculosis is a global public health problem, especially in emerging countries. Mycobacterium tuberculosis is the main cause of cervical lymphadenopathy; nontuberculous mycobacteria are relatively common in children and rare in adults. Objective: To identify and establish the frequency of infectious etiology by nontuberculous mycobacteria in Mexican adult patients with cervical lymphadenopathy. Methods: The study included 85 patients over 18 years with cervical lymphadenopathy; 45 were HIV-positive, 40 were HIV-negative; they had no history of tuberculosis treatment and were selected from a third-level hospital. It was carried out a biopsy of the lymph node for the histopathological study, a search for acid-fast bacilli, a tube culture to indicate growth of Mycobacterium BACTEC (MGIT-960) and identification of mycobacterial strain by PCR-RFLP (restriction fragment length polymorfism) of hsp65. Results: In 42 HIV-positive patients (93%), strains corresponded to Mycobacterium tuberculosis complex, two (4.4%) to M. intracellulare and one (2.2%) to M. gordonae. Among HIV-negative patients, 39 of strains (97.5%) corresponded to patients with M. tuberculosis complex and one strain (2.5%) to M. fortuitum. Conclusion: The presence of nontuberculous mycobacteria was found in 4.7% of all cases. Despite this low frequency, it must be taken into account as a possible cause of lymphadenopathy, since its prompt identification enables introducing specific treatment.

Introducción: la tuberculosis es un problema de salud pública mundial, sobre todo en países emergentes. El Mycobacterium tuberculosis es el principal causante de las adenopatías cervicales; las micobacterias no tuberculosas son relativamente frecuentes en el niño y raras en adultos. Objetivo: identificar y establecer la frecuencia de la etiología infecciosa por micobacterias no tuberculosas (MNT) en pacientes adultos mexicanos con linfadenopatias cervicales. Métodos: se estudiaron 85 pacientes mayores de 18 años, con linfadenopatía cervical, 45 con positividad al virus de la inmunodeficiencia humana (VIH) y 40 VIH negativos, sin antecedentes de tratamiento antituberculoso, seleccionados en un hospital de concentración de especialidad de tercer nivel. Se realizó biopsia de nodo linfático para su estudio histopatológico, búsqueda de bacilos ácido-alcohol resistentes, cultivo en el tubo indicador del crecimiento de Mycobacterium BACTEC (MGIT-960) y la identificación de cepa micobacteriana por PCR-RFLP (restriction fragment lenght polymorfism) de hsp65. Resultados: las cepas correspondieron al complejo Mycobacterium tuberculosis en 42 pacientes VIH positivos (93%), dos (4.4%) a M. intracellulare y una (2.2%) a M. gordonae. Las cepas correspondieron al complejo M. tuberculosis en 39 pacientes VIH negativos (97.5%) y una a M. fortuitum (2.5%). Conclusión: la presencia de MNT se encontró en 4.7% de todos los casos. A pesar de su baja frecuencia, deben ser tomadas en cuenta como posible causa de linfadenopatías, porque su identificación oportuna permite instaurar un tratamiento específico.

Identification of risk factors and diagnostic methodology in serositis by M. tuberculosis / Identificación de factores de riesgo y metodología diagnóstica en serositis por M. tuberculosis

Background: Tuberculosis is a public health problem, extrapulmonary presentations have increased, it is difficult to diagnose because of the low bacillary load. Objective: To identify risk factors and to evaluate the efficiency of diagnostic methods in pleural, meningeal, peritoneal and pericardial tuberculosis. Methods: Prospective study of cases and controls. A multiple conditional logistic regression model was used to identify risk factors. Biopsy was performed and 7 mL of fluid was extracted from the affected site, Löwestein-Jensen and MGITI960 culture, Ziehl-Neelsen staining, adenosine deaminase and endpoint PCR directed to the insertion sequence 1S6110 for M. tuberculosis were performed. Results: 116 patients were included, in 58 M. tuberculosis was confirmed by positive culture (meningeal Tb 34 cases, pleural 14, peritoneal 8, pericardial 2 cases) and 58 serositis of non-tuberculous etiology. Being a carrier of HIV and living with people infected with tuberculosis were the main risk factors OR = 3.6 and OR = 6.8. The staining had sensitivity of 25.9%, PCR of 65.5% and adenosine deaminase with 82.8% Conclusions: Conventional diagnostic methods had low efficacy, adenosine deaminase and molecular biology techniques are the most useful, in our environment these tests should be performed immediately in patients with risk factors and suspected serositis of tuberculous origin.

Introducción: la tuberculosis es un problema de salud pública, las presentaciones extrapulmonares han aumentado, siendo de difícil diagnóstico por su baja carga bacilar. Objetivo: identificar los factores de riesgo y evaluar la eficacia de los métodos diagnósticos en la tuberculosis pleural, meníngea, peritoneal y pericárdica. Métodos: estudio prospectivo de casos y controles. Se empleó un modelo de regresión logística condicional múltiple para identificar factores de riesgo. Se realizó biopsia y se extrajeron siete mL de líquido presente del sitio afectado, se realizó cultivo Löwestein-Jensen y MGITI960, tinción Ziehl-Neelsen, adenosina deaminasa y PCR en punto final dirigida a la secuencia de inserción 1S6110 para M. tuberculosis. Resultados: se incluyeron 116 pacientes, en 58 se confirmó M. tuberculosis por cultivo positivo (Tb meníngea 34 casos, pleural 14, peritoneal 8, pericárdica 2 casos) y 58 serositis de etiología no tuberculosa. Ser portador de VIH y convivir con personas infectadas con tuberculosis fueron los mayores factores de riesgo OR = 3.6 y OR = 6.8. La tinción tuvo sensibilidad de 25.9%, PCR de 65.5% y adenosina deaminasa con 82.8%. Conclusiones: los métodos diagnósticos convencionales tuvieron baja eficacia, la adenosina deaminasa y las técnicas de biología molecular son los de mayor utilidad, en nuestro medio estos estudios deben realizarse de inmediato en pacientes con factores de riesgo y sospecha de serositis de origen tuberculoso.

Nontuberculous mycobacteria in clinical samples with negative acid-fast bacilli.

BACKGROUND: There is a progressive increase in nontuberculous mycobacteria (NTM) in pulmonary and extrapulmonary infections that might cause confusion with the Mycobacterium tuberculosis complex. To determine the frequency of finding NTM in clinical samples from patients diagnosed with active tuberculosis, with negative acid-alcohol-resistant bacilli (acid-fast bacillus [AFB]) in a third-level specialty hospital's mycobacterial laboratory between January 2013 and December 2014. METHODS: This is a prospective, descriptive study where isolated strains of biological material were studied in Lowenstein-Jensen and BACTEC MGIT 960 cultures. RESULTS: Clinical samples of 120 patients were studied, with pulmonary samples of 99/120 (82%) and extrapulmonary samples of 21/120 (18%). We identified NTM in 37/120 samples (30.8%), of which 16 in pulmonary, 13 in genitourinary, 3 in bone marrow, and 5 in various specimens. Mycobacterium avium was isolated in 20 samples, Mycobacterium intracellulare in seven samples, and various other species of NTM in the other 10 samples. CONCLUSION: To establish adequate treatment, we point out the importance of identifying the presence of NTM in the clinical samples of active tuberculosis patients with negative AFB, as possibly becoming confused with M. tuberculosis and which is essential in deciding which treatment is the most adequate.

Skin biopsy: a pillar in the identification of cutaneous Mycobacterium tuberculosis infection.

INTRODUCTION: The present study aimed to establish the frequency and clinical characteristics of cutaneous tuberculosis among Mexican adult patients. METHODOLOGY: Ninety-five patients with clinically compatible lesions to cutaneous tuberculosis participated in the study. All patients were HIV negative and none of them had previous anti-TB treatment. A skin biopsy was taken from every patient suspected of having tuberculosis, and a histopathologic examination was performed as follows: Ziehl-Neelsen staining; culturing of mycobacteria by Löwenstein-Jensen (L-J) medium; Mycobacteria Growth Indicator Tube detection via BACTEC (MGIT-360); and polymerase chain reaction (PCR) with the sequence of insertion IS6110 for Mycobacterium tuberculosis complex. RESULTS: Tuberculosis was confirmed in 65 out of 95 cases (68.4%). Identified lesions were scrofuloderma (42 cases, 64.6%); lupus vulgaris (12 cases, 18.4%); warty tuberculosis (six cases, 9.2%); and papulonecrotic tuberculoid (five cases; 7.7%). The Ziehl-Neelsen staining was positive for acid fast bacilli in nine cases (13.8%) and 48 patients were positive for the PCR amplification (73.8%). All skin biopsies resulted positive for tuberculosis. A positive clinical response to the specific treatment was considered a confirmation for tuberculosis. The noninfectious etiology corresponded to 30 cases (31.6%). CONCLUSIONS: Tuberculosis in developing countries is still an important cause of skin lesions which must be studied via histopathological examination and culture due to their low bacillary load. A PCR test is necessary to obtain faster confirmation of the disease and to establish an early, specific and effective treatment.