Objective: To determine the phenotypic and genotypic features of extended spectrum beta-lactamase (ESBL) producing strains of Escherichia coli isolated from urine samples of patients attending outpatient services in a private laboratory in Lima, Peru. Methods: 53 E. coli isolates were evaluated using two phenotypic methods: Jarlier and CLSI, the susceptibility profile was performed using the disk diffusion method and the genotypic features were analyzed using PCR for detecting blaCTX-M, blaTEM y blaSHV genes. Results: The 53 ESBL producing strains of E. coli accounted for 16,30% of all E. coli isolates affecting mostly women older than 65 years. High resistant profile to AMP, CEF, CRO (100%), LEV (87%), NOR (92%), CIP, NAL (94%), CXM, CTX (96%), SXT (70%), ATM (75%) and TOB (85%) was observed. High susceptibility to NIT, IPM (100%), AMK (91%) and FOF (73.6%) was observed. The most frequent bla gen was blaCTX-M (55%), followed by blaCTX-M+TEM (24%), blaTEM (13%) and blaSHV (6%). Conclusions: The rate of ESBL producing strains of E. coli was 16.3% and the blaCTX-M gen was the most common gene type. These results provide valuable information for starting empiric antibiotic therapy in this setting.

Objetivos: Determinar las características fenotípicas y genotípicas de las β-lactamasas de espectro extendido (BLEE) en E. coli aislados de cultivos de orina de pacientes de la comunidad en un laboratorio privado de la ciudad de Lima, Perú. Material y métodos: Se evaluaron 53 aislamientos de E. coli por dos métodos fenotípicos: Jarlier y CLSI, el perfil de susceptibilidad se realizó mediante disco difusión y la caracterización genotípica mediante PCR para los genes blaCTX-M, blaTEM y blaSHV. Resultados: Los 53 aislamientos productores de BLEE representaron el 16,30% del total de aislados de E. coli, afectando principalmente a mujeres mayores de 65 años. El perfil de susceptibilidad evidenció alta resistencia a AMP,CEF,CRO(100%), LEV(87%), NOR(92%), CIP y NAL(94%), CXM y CTX(96%),SXT(70%), ATM(75%) y TOB (85%); asimismo elevada sensibilidad a NIT e IPM(100%), AMK(91%) y FOF(73,6%). El tipo de gen bla más frecuente fue blaCTX-M (55%), seguido por la coexistencia blaCTX-M+TEM (24%), blaTEM (13%) y blaSHV (6%). Conclusiones: La frecuencia de E. coli productores de BLEE fue de 16,3%; siendo el gen tipo blaCTX-M el más frecuente, información valiosa para orientar la terapia antimicrobiana empírica.

Conducting a same day smears is a strategy that has recently been adopted by the World Health Organization. Our study sought to determine the additional diagnostic yield of a second sputum smear taken on the same day among patients with respiratory symptoms, attended at the emergency room from two hospitals in Lima. We included 270 patients from January to November 2011, which were asked two samples of sputum within two hours; the samples were processed by the Ziehl Neelsen method. The frequency of positive sputum smears was 18.5%. The additional diagnostic yield for the second smear was 20.9%. Considering the high prevalence of tuberculosis diagnosed by sputum smear, it is suggested that this test be included routinely in the emergency rooms of hospitals in Lima Metro area.

La realización de una baciloscopía el mismo día de la atención del paciente es una estrategia que recientemente ha sido adoptada por la Organización Mundial de la Salud. Nuestro estudio buscó determinar el rendimiento diagnóstico adicional de una segunda baciloscopía tomada en un mismo día en pacientes con síntomas respiratorios, atendidos en el servicio de emergencia de dos hospitales de Lima. Se incluyeron 270 pacientes desde enero a noviembre de 2011, a los cuales se les solicitó dos muestras de esputo con un lapso de dos horas; las muestras fueron procesadas mediante el método de Ziehl Neelsen. La frecuencia de baciloscopías positivas fue del 18,5%. El rendimiento diagnóstico adicional de la segunda baciloscopía fue 20,9%. Considerando la alta frecuencia de tuberculosis diagnosticada por baciloscopía, se sugiere que esta prueba sea incluida de manera rutinaria en las emergencias de los hospitales de Lima Metropolitana.

Conducting a same day smears is a strategy that has recently been adopted by the World Health Organization. Our study sought to determine the additional diagnostic yield of a second sputum smear taken on the same day among patients with respiratory symptoms, attended at the emergency room from two hospitals in Lima. We included 270 patients from January to November 2011, which were asked two samples of sputum within two hours; the samples were processed by the Ziehl Neelsen method. The frequency of positive sputum smears was 18.5%. The additional diagnostic yield for the second smear was 20.9%. Considering the high prevalence of tuberculosis diagnosed by sputum smear, it is suggested that this test be included routinely in the emergency rooms of hospitals in Lima Metro area

La realización de una baciloscopía el mismo día de la atención del paciente es una estrategia que recientemente ha sido adoptada por la Organización Mundial de la Salud. Nuestro estudio buscó determinar el rendimiento diagnóstico adicional de una segunda baciloscopía tomada en un mismo día en pacientes con síntomas respiratorios, atendidos en el servicio de emergencia de dos hospitales de Lima. Se incluyeron 270 pacientes desde enero a noviembre de 2011, a los cuales se les solicitó dos muestras de esputo con un lapso de dos horas; las muestras fueron procesadas mediante el método de Ziehl Neelsen. La frecuencia de baciloscopías positivas fue del 18,5%. El rendimiento diagnóstico adicional de la segunda baciloscopía fue 20,9%. Considerando la alta frecuencia de tuberculosis diagnosticada por baciloscopía, se sugiere que esta prueba sea incluida de manera rutinaria en las emergencias de los hospitales de Lima Metropolitana

Background : Staphylococcus aureus is an important pathogen involved in a series of infections and toxin mediated syndromes, has many virulence factors and its impact increases its resistance to antimicrobial agents. Objectives: To determine the frequency of community acquired methicillin resistant Staphylococcus aureus among hospitals in Lima -Peru. Material and Methods: We performed a prospective multicenter study. The resistance to methicillin was determined by the Oxacillin Agar Screen method. The origin of the strain was determined using CDC criteria, the Panton Valentine Leucocidin was identified by molecular methods. Results: We isolated 276 strains of Staphylococcus aureus, 160 were resistant to methicillin (58%). 9 strains were identified as community acquired MRSA (5.6%). The Panton Valentine Leucocidin was identified in 25 strains (9.1%), 14 were MSSA and 11 MRSA, only 4 of the latter were CA MRSA, 7 were HA MRSA (p <0.001). Conclusions: The study showed frequencys of methicillin resistance, but low CA MRSA. In our environment the presence of PVL would not be a marker for the identification of CA MRSA. (Rev Med Hered 2010;21:4-10).

Staphylococcus aureus es un importante patógeno involucrado en una serie de infecciones e intoxicaciones, presenta múltiples factores de virulencia y su impacto se incrementa por su notable resistencia a los antimicrobianos. Objetivo: Determinar la frecuencia de Staphylococcus aureus meticilino resistente adquiridos en la comunidad, en hospitales de Lima- Perú. Material y métodos: Se realizó un estudio descriptivo multicéntrico. La resistencia a meticilina se determinó por el método Oxacillin Agar Screen. El origen de la cepa fue determinado mediante los criterios de los CDC; la Leucocidina de Panton Valentine fue identificada por métodos moleculares. Resultados: Se aislaron 276 cepas de Staphylococcus aureus, 160 fueron resistentes a meticilina (58%), 9 de ellas fueron identificadas como adquiridas en la comunidad (5,6%). La PVL fue identificada en 25 cepas (9,1%), 14 fueron MSSA y 11 MRSA, de éstas últimas solo 4 fueron MRSAcom, 7 fueron MRSAhosp (p<0,001). Conclusiones: El estudio revela niveles elevados de resistencia a meticilina, pero niveles bajos de MRSAcom. En nuestro medio la presencia de PVL no constituiría un marcador para la identificación de los MRSAcom. (Rev Med Hered 2010;21:4-10).

Objective. To evaluate the diagnostic yield of the media Middlebrook 7H11 combined with human or ovine blood in comparison with the Ogawa solid media for the diagnosis of pulmonary tuberculosis. Material and methods. We evaluated sputum samples of patients with clinical suspicion of pulmonary tuberculosis. The samples were seeded in Middlebrook 7H11 agar associated with human or ovine blood and in Ogawa media. Results. A total of 130 samples were collected. The positivity for M.tuberculosis in Middlebrook 7H11/human blood, Middlebrook 7H11/sheep blood and Ogawa was 45.38%, 46.15% and 43.84% respectively. The mean times for growth for the M. tuberculosis were 12.81± 6.52, 13.07 ± 6.63 and 20.14 ± 6.75 days (p< 0,01 for the comparison of agar based medium versus Ogawa). Conclusions. The combination of Middlebrook 7H11 with sheep or human blood has a comparable diagnostic yield but a shorter growth time than the Ogawa solid media.

Objetivo. Evaluar el rendimiento diagnóstico del medio Middlebrook 7H11 combinado con sangre humana u ovina en comparación con el medio sólido Ogawa para el diagnóstico de tuberculosis pulmonar. Materiales y métodos. Se evaluaron muestras de esputo provenientes de pacientes con sospecha de tuberculosis pulmonar. Las muestras fueron sembradas en agar Middlebrook 7H11 asociado a sangre humana u ovina y en medio Ogawa. Resultados. Se recolectaron un total de 130 muestras. La positividad para M.tuberculosis en Middlebrook 7H11/sangre humana, Midlebrook 7H11/sangre ovina y Ogawa fue de 45,38%, 46,15% y 43,84% respectivamente. El tiempo de crecimiento promedio del Mycobacterium tuberculosis fue de 12,81 ± 6,52, 13,07 ± 6,63 y 20,14 ± 6,75 días (p<0,01 para la comparación de los medios basados en agar vs Ogawa). Conclusiones. La combinación de Middlebrook 7H11 con agar sangre u ovina presenta un rendimiento diagnóstico comparable y un tiempo de crecimiento menor que el medio sólido Ogawa.

This study developed a clinical score based on clinical and radiographic data for the diagnosis of smear-negative pulmonary tuberculosis (SNPT). SNPT was defined as a positive culture in Ogawa in a patient with two negative sputum smears. Data from patients admitted to the emergency ward with respiratory symptoms and negative acid-fast bacilli (AFB) smears was analyzed by means of logistic regression to develop the predictive score.Two hundred and sixty two patients were included. Twenty patients had SNPT. The variables included in the final model were hemoptysis, weight loss, age > 45 years old, productive cough, upper-lobe infiltrate, and miliary infiltrate. With those, a score was constructed. The score values ranged from -2 to 6. The area under the curve for the ROC curve was 0.83 (95% CI 0.74-0.90). A score of value 0 or less was associated with a sensitivity of 93% and a score of more than 4 points was associated with a specificity of 92% for SNPT. Fifty-two point twenty-nine percent of patients had scores of less than one or more than four, what provided strong evidence against and in favor, respectively, for the diagnosis of SNPT. The score developed is a cheap and useful clinical tool for the diagnosis of SNPT and can be used to help therapeutic decisions in patients with suspicion of having SNPT.

Background: Rifampicin resistance in M. tuberculosis involves mutations in the rpoβ gene, encoding the β subunit of RNA polymerase. Objective: To identify the rpoβ gene mutations in strains of M. tuberculosis isolated from the Health Sub-Region in North Lima, Perϊ. Materials and methods: 73 sputum samples from patients with pulmonary tuberculosis were cultured on Lφwenstein-Jensen media. 62 samples, with more than 10 colonies per tube, were analyzed for susceptibility to isoniazid, streptomycin, rifampicin and ethambutol. DNA was isolated, amplified by PCR, followed by the cloning of the amplified product in the vector pGEM-T, transformation of competent E. coli bacteria, selection of recombinant clones and sequencing of the plasmid DNA for the determination of the polymorphisms in the rpoβ gene. Results: 52 strains out of 73 isolates (83,9%) were resistant to rifampicin (Rif r ), while 10 isolates (16,1%) were found to be rifampicin susceptible (Rif s ). 51 out of 52 Rif r strains contained mutations in the rpoβ gene associated with rifampicin resistance. 20 mutations were identified. The most frequently observed mutations were localized in the codons Ser-531 (62,7 %), His-526 (15,72%), Asp-516 (11,78%) and Gln-513 (5,88 %). No mutations were detected in the 10 Rif s strains. 94,2% of the Rif r strains were also resistant to isoniazid. Conclusions: rpoβ gene mutations associated with rifampicin resistance were present in almost all Rif r strains analyzed. Almost all Rif r strains were also resistant to isoniazid.

Antecedentes: La resistencia a rifampicina en M. tuberculosis involucra mutaciones en el gen rpoβ que codifica a la subunidad β de la ARN polimerasa. Objetivo: Identificar las mutaciones del gen rpoβ , en cepas de M. tuberculosis asociadas con resistencia a rifampicina aisladas de la Subregiσn de Salud Lima Norte, Perϊ. Materiales y métodos: Se cultivó en Lowestein - Jenseen 73 muestras de esputo de pacientes con tuberculosis pulmonar. A 62, con más de 10 colonias por tubo, se les comprobó susceptibilidad a isoniazida, rifampicina, estreptomicina y etambutol. Se realizó la extracción de ADN por PCR, clonación en el vector pGEM-T, transformación, selección de clonas recombinantes y secuenciamiento del ADN plasmídico para la determinación de los polimorfismos del gen rpoβ. Resultados: 52 (83,9%) cepas fueron resistentes a rifampicina (Rif r) y 10 (16,1%) susceptibles (Rif s ). Se encontró alteraciones en el gen rpoβ en 51 de 52 cepas Rif r Se identificaron 20 mutaciones. Las mutaciones mαs frecuentes fueron encontradas en los codones Ser-531 (62,7%), His-526 (15,7%), Asp-516 (11,8%) y Gln-513 (5,9%). No se observσ mutaciσn alguna en las 10 cepas Rif s. 94,2% de nuestras cepas Rif r fueron también resistentes a isoniazida. Conclusiones: Se encontraron mutaciones en el gen rpoB de casi todas las cepas Rif r ; asimismo, casi todas las cepas Rif r fueron también resistentes a isoniazida.