Caracterización fenotípica y molecular de cepas de Klebsiella pneumoniae productores de carbapenemasas tipo OXA-48 circulantes en Chile

Introducción: La aparición y diseminación de Enterobacterales resistentes a carbapenémicos ha generado un gran impacto en las infecciones asociadas a la atención de salud en el mundo. Recientemente, en Chile se detectó un brote por Klebsiella pneumoniae productora de carbapenemasas tipo oxacilinasas (OXA) de la subfamilia tipo OXA-48, reportándose los primeros casos en pacientes hospitalizados mayoritariamente en la zona norte del país. Objetivo: Determinar los perfiles fenotípicos, genotípicos y de susceptibilidad antimicrobiana de 16 cepas referidas durante mayo del año 2021 desde las regiones de Antofagasta y Metropolitana al Laboratorio de Referencia del Instituto de Salud Pública. Metodología: Las cepas provenientes de muestras clínicas fueron analizadas mediante técnicas tradicionales (Kirby-Bauer y epsilometría) y automatizadas, además de técnicas colorimétricas, inmunocromatográficas y moleculares (RPC y PFGE). Resultados: Se detectó la presencia de los genes blaoxa-48 y blaoxa-232 con una resistencia inusual, tanto a carbapenémicos (ertapenem, imipenem y meropenem) como a cefalosporinas (cefepime, cefotaxima y ceftazidima), además de piperacilina/tazobactam y temocilina. Se detectaron dos subtipos por PFGE, siendo predominante el clon CL-Kpn-Spe-329 (93,8%) con dos mecanismos de resistencia identificados: carbapenemasa y β-lactamasa de espectro extendido (BLEE). Conclusión: Ante esta alerta epidemiológica es necesario unificar criterios existentes en la red asistencial nacional para la oportuna detección, vigilancia y control de posibles brotes de cepas productores de oxacilinasa tipo OXA-48.

Background: The appearance and spread of carbapenems-resistant Enterobacterales have generated a major impact on health care-associated infections worldwide. Recently, a Klebsiella pneumoniae outbreak expressing OXA-48 like-carbapenemases was detected in Chile, the first reported cases corresponded to hospitalized patients mainly from northern Chile. Aim: To characterize the phenotypic and genotypic profiles of antimicrobial susceptibility of 16 clinical isolates referred during May 2021 from Antofagasta and Metropolitan regions to the Reference Laboratory of Instituto de Salud Publica. Methods: Antimicrobial susceptibility of all strains was analyzed using traditional (Kirby-Bauer and epsilometry) and automated methods, and complemented with colorimetric, immunochromatographic and molecular (PCR and PFGE) techniques. Results: As a result of the genetic characterization, blaoxa-48 and blaoxa-232 genes were detected, showing the isolates an unusual resistance profile to both carbapenems (ertapenem, imipenem, and meropenem) and cephalosporins (cefepime, cefotaxime, and ceftazidine), as well as piperacillin/ tazobactam and temocillin. Two subtypes were detected by PFGE, with a predominant clone CL-Kpn-Spe-329 (93.8%), with two resistance mechanisms identified: carbapenemase and extended-spectrum β-lactamase (ESBL). Conclusion: Due to this epidemiological alert, it is essential the establishment of national guidelines for early detection, surveillance, and control of future outbreaks of OXA-48 like carbapenemases isolates.

Enterobacterales productores de carbapenemasas en sifones de lavamanos de una Unidad de Paciente Crítico

Resumen Introducción: Enterobacterales productores de carbapenemasas (EPC) son una importante causa de infecciones asociadas a la atención en salud (IAAS). El principal reservorio de EPC lo constituyen pacientes infectados y colonizados, sin embargo, también se han identificado reservorios ambientales. Objetivo: Detectar la presencia de EPC en los sifones de lavamanos de la unidad de cuidados críticos de pacientes quemados adultos (UPC QMD) y unidad de cuidados críticos de pacientes pediátricos crónicos (UCEP). Método: Se recolectaron cuatro muestras de sifones de los lavamanos ubicados en el interior de las unidades de pacientes en UCEP y 10 de UPC QMD. A las muestras se les realizó estudio fenotípico y molecular para detección de carbapenemasas en el Instituto de Salud Pública de Chile. Resultados: En los sifones estudiados de UCEP no se aislaron cepas de EPC. En UPC QMD, 50% de los sifones estudiados se aislaron cepas de EPC. Conclusiones: En UPC QMD se objetivó la presencia de EPC en una alta proporción de los sifones de lavamanos testeados, lo que demuestra un reservorio ambiental de bacterias multi-resistentes.

Abstract Introduction: Carbapenemase-producing Enterobacterales (CPE) are an important cause of health care associated infections (HAI). The main reservoir is constituted by infected and colonized patients; however, environmental reservoirs have also been identified. Objective: To detect the presence of CPE in the sink traps of the critical care unit for adult burn patients (UPC QMD) and the critical care unit for chronic pediatric patients (UCEP). Material and Method: Four samples of trap were collected from the sinks located inside the patient units at PICU and 10 at UPC QMD. The samples underwent a phenotypic and molecular study for the detection of carbapenemases at the Institute of Public Health of Chile. Results: In the UCEP no EPC strains were isolated. In UPC QMD, CPE was detected in 50% of the traps. Conclusions: In UPC QMD, the presence of CPE was observed in a high proportion of the tested sinks traps, which shows an environmental reservoir of multi-resistant bacteria.

Calidad de vida en personas mayores de entornos rurales: diferencias entre hombres y mujeres

RESUMEN Objetivo: Analizar las diferencias entre hombres y mujeres en los diversos dominios de la calidad de vida (CdV) en personas mayores que residen en zonas rurales del extremo norte de Chile. Material y Método: Estudio cuantitativo, descriptivo, transversal realizado entre noviembre 2019 y febrero 2020 en diversas zonas rurales de la Región de Arica y Parinacota. Contó con un universo de 100 personas mayores de 60 años y sin deterioro cognitivo. Se aplicó el Cuestionario Breve de Calidad de Vida (CUBRECAVI). La investigación fue aprobada por la Escuela de Trabajo Social de la Universidad de Tarapacá. Resultados: 69% mujeres, 63% entre 60 y 69 años, 67% casado o en pareja y 58% indígena. Se confirman diferencias estadísticamente significativas en el dominio salud objetiva: tobillos hinchados, cansancio, dificultad para dormir, escape de orina y hormigueos; en el dominio salud psíquica: llorar con facilidad, sentimientos depresivos y problemas de memoria. Lo mismo se observa en integración social, en el nivel de satisfacción con la relación del cónyuge o pareja, con los familiares y con las amistades. Igualmente los hallazgos confirman diferencias en el dominio actividades, nivel de actividades, frecuencia camina y frecuencia hace manualidades. Por último, en el dominio calidad ambiental, en la satisfacción con la temperatura del hogar. Los resultados indican que las mujeres tienen peor CdV y las personas mayores perciben que el dominio salud es el más importante para valorar su CdV. Conclusión: Se confirma riesgo biopsicosocial para mujeres mayores que residen en zonas rurales del extremo norte de Chile. Enfermería y otras disciplinas de trato directo con las personas mayores deben conocer las distintas maneras de envejecer, diferenciando por sexo y especificidades del entorno rural.

ABSTRACT Objective: To analyze the differences between men and women in the different domains of quality of life (QoL) among elderly people living in rural areas of the north of Chile. Material and Method: Quantitative, descriptive, cross-sectional study conducted between November 2019 and February 2020 in various rural areas of the Region of Arica and Parinacota. The sample consisted of 100 people over 60 years old and without cognitive impairment. The Brief Quality of Life Questionnaire (CUBRECAVI) was applied. The research was approved by the School of Social Work of the Universidad de Tarapacá. Results: 69% were women, 63% were between 60 and 69 years old, 67% were married or had a relationship and 58% had indigenous origin. Statistically significant differences were confirmed in the objective health domain: swollen ankles, tiredness, sleep problems, urinary incontinence and tingling; in the psychic health domain: crying easily, depressive feelings and memory problems. This was similar in the field of social integration, in the level of satisfaction concerning the relationship with the spouse or partner, with family members and with friends. Moreover, the findings corroborate differences in the domain of activities, level of activities, frequency of walking and frequency of doing crafts. Finally, in the environmental quality domain, there are differences in satisfaction with room temperature. The results show that women have worse QoL and the elderly perceive that the health domain is the most important when assessing their QoL. Conclusion: There is a biopsychosocial risk for older women living in rural areas in the north of Chile. It is clear that nursing and other areas dealing directly with the elderly have to familiar with the different ways of aging, taking into account gender and specific rural setting.

RESUMO Objetivo: Analisar as diferenças entre homens e mulheres nos diversos domínios da qualidade de vida (QV) em idosos residentes em áreas rurais do norte do Chile. Material e Método: Estudo quantitativo, descritivo, transversal realizado entre novembro de 2019 e fevereiro de 2020 em diversas áreas rurais da Região de Arica e Parinacota. A amostra consistiu em 100 pessoas com mais de 60 anos e sem déficit cognitivo. Foi aplicado o Questionário Breve de Qualidade de Vida (CUBRECAVI). A pesquisa foi aprovada pela Escola de Serviço Social da Universidade de Tarapacá. Resultados: 69% eran mulheres, 63% tinham entre 60 e 69 anos, 67% estavam casadas ou tinham um relacionamento e 58% tinham origem indígena. Foram confirmadas diferenças estatisticamente significativas no dominio da saúde objetiva: tornozelos inchados, fadiga, dificuldade para dormir, perda de urina e formigamento; no domínio da saúde psíquica: choro com facilidade, sentimentos depressivos e problemas de memória. O mesmo foi observado na integração social, no nível de satisfação na relação com o cônjuge ou companheiro, com os familiares e com os amigos. Da mesma forma, os achados confirmam diferenças no domínio das atividades, nível de atividades, frequência de caminhadas e frequência de artesanato. Finalmente, no domínio da qualidade ambiental há diferenças no grau de satisfação com a temperatura da casa. Os resultados indicam que as mulheres apresentam pior QV e os idosos percebem que o domínio saúde é o mais importante para avaliar sua QV. Conclusão: O risco biopsicossocial é confirmado para mulheres idosas residentes em áreas rurais do extremo norte do Chile. A enfermagem e as demais disciplinas que lidam diretamente com o idoso devem conhecer as diferentes formas de envelhecer, diferenciando entre os sexos e as especificidades do meio rural.

Caracterización genética de cepas de Listeria monocytogenes aisladas durante los años 2007-2014 en Chile / Genetic characterization of Listeria monocytogenes strains isolated during 2007-2014 in Chile

Resumen Introducción: Listeria monocytogenes es un patógeno transmitido por alimentos que causa listeriosis, una enfermedad que puede presentarse como gastroenteritis febril o en una forma invasora que tiene altas tasas de mortalidad. Hasta el momento, ha sido poco estudiada la diversidad genética de cepas de L. monocytogenes aisladas desde pacientes, alimentos y fuentes ambientales en Chile. Objetivo: Caracterizar genéticamente cepas de L. monocytogenes de estos tres orígenes recibidas por el Instituto de Salud Pública de Chile (ISP) entre los años 2007 y 2014. Material y Métodos: Se seleccionaron 94 cepas de L. monocytogenes correspondientes a 94 pulsotipos diferentes identificados por electroforesis en gel de campo pulsado (PFGE), se extrajo ADN y se realizó serotipificación mediante reacción de polimerasa en cadena (RPC) y tipificación de secuencias multilocus (MLST). Resultados: El serotipo más común fue 4b (55,3%), seguido de 1/2a (25,5%), 1/2b (17%) y 1/2c (2,2%). Se identificaron 32 secuencias tipo (ST), de las cuales cuatro fueron nuevas, y las predominantes fueron ST1 (28,7%) y ST2 (13,8%). La totalidad de las cepas se agrupó en los Linajes I y II. Conclusiones: Se observó una gran variabilidad genética en las cepas de L. monocytogenes analizadas, siendo predominantes las secuencias tipo ST1 y ST2, ambas pertenecientes al Linaje I. Nuestros resultados contribuyen a conocer la estructura poblacional de este patógeno en Chile y su presencia en muestras clínicas, alimentos y el medio ambiente.

Background: Listeria monocytogenes is a foodborne pathogen that causes listeriosis, a disease that can present as febrile gastroenteritis or as an invasive form that has high mortality rates. So far, the genetic diversity of strains of L. monocytogenes isolated from patients, foods and environmental sources in Chile has been poorly studied. Aim: To characterize genetically L. monocytogenes strains received by the Institute of Public Health of Chile (ISP) between 2007 and 2014. Methods: We selected 94 strains of L. monocytogenes corresponding to 94 different pulsotypes identified by pulsed field gel electrophoresis (PFGE), DNA was extracted and serotyping was performed by polymerase chain reaction (PCR) and multilocus sequence typing (MLST). Results: The most common serotype was 4b (55.3%), followed by serotypes 1/2a (25.5%), 1/2b (17%) and 1/2c (2.2%). 32 sequence-type (ST) were identified, of which 4 were new, and the predominant ones were ST1 (28.7%) and ST2 (13.8%). All the strains of L. monocytogenes were grouped in Lineages I and II. Conclusions: A great genetic variability was observed in the strains of L. monocytogenes analyzed, being predominant the ST1 and ST2, both belonging to Lineage I. Our results contribute to know the population structure of this pathogen in Chile and its presence in clinical samples, food and the environment.

Novel nonsense mutation in the katA gene of a catalase-negative Staphylococcus aureus strain

Abstract We report the first description of a rare catalase-negative strain of Staphylococcus aureus in Chile. This new variant was isolated from blood and synovial tissue samples of a pediatric patient. Sequencing analysis revealed that this catalase-negative strain is related to ST10 strain, which has earlier been described in relation to S. aureus carriers. Interestingly, sequence analysis of the catalase gene katA revealed presence of a novel nonsense mutation that causes premature translational truncation of the C-terminus of the enzyme leading to a loss of 222 amino acids. Our study suggests that loss of catalase activity in this rare catalase-negative Chilean strain is due to this novel nonsense mutation in the katA gene, which truncates the enzyme to just 283 amino acids.

Staphylococcus aureus resistente a meticilina asociado a la comunidad (SARM-AC): comunicación de los primeros cuatro casos pediátricos descritos en Hospital de Niños Roberto del Río / Community associated-methicillin-resistant Staphylococcus aureus (SAMR-AC): comunication of the first four pediatric cases in the Roberto del Rio Children's Hospital

Staphylococcus aureus is a known pathogen in pediatric patients that produces skin infections, cutaneous abscess, cellulitis and osteoarticular infections. Most of these infections are produced by a meticilin susceptible strain. The community associated methicillin resistant Staphylococcus aureus was published for the first time in 1993, ever since then is has been recognized as a cosmopolite pathogen. The first report in Latin America was published in 2003, and in Chile in 2008 from adult patients that have reported traveling to other countries. The following series describes four pediatric cases, all school-aged children, diagnosed since 2012 with clinical followups and molecular studies. Two cases presented as osteomyelitis of the lower extremity; and one presented as arm cellulitis. These three cases had Panton Valentine leukocidine (PV-L) negative strains from the clone complex 8. The last case presented a renal abscess, the strain was PV-L positive from the clone complex 30. This case series constitutes the first pediatric case report in Chile.

Staphylococcus aureus es un patógeno conocido como causa de infecciones de piel, tejidos blandos, osteoarticulares y celulitis en niños. Estas infecciones son principalmente causadas por cepas sensibles a meticilina. Staphylococcus aureus resistente a meticilina asociado a la comunidad fue publicado por primera vez en población australiana nativa en 1993 y desde entonces se ha transformado en un patógeno cosmopolita. En Latinoamérica se publicó el primer caso en 2003 y en Chile se comunicaron los primeros casos en 2008, casi todos adultos y con antecedentes de viaje al extranjero. Nuestra serie describe cuatro casos clínicos en niños escolares, pesquisados desde el 2012 con seguimiento clínico y estudio molecular. Dos casos se presentaron como una osteomielitis de extremidad inferior y uno como una celulitis de brazo. Los tres primeros casos correspondieron a cepas leucocidina Panton Valentine (PV-L) negativa, del complejo clonal 8. El último caso fue un absceso renal, cepa PV-L positiva, perteneciente al complejo clonal 30. Esta serie es el primer reporte de casos pediátricos en Chile.

Vigilancia de laboratorio de Streptococcus pneumoniae procedente de enfermedad invasora, Chile 2007-2012 / Laboratory surveillance of Streptococcus pneumoniae from invasive disease, Chile 2007-2012

Background: 10-valent pneumococcal vaccine (PCV-10) was introduced in 2011 to the National Immunization Program in Chile. It was administered in 4 doses, but in 2012 it was modified to a 3 dose program. This article shows the results of the Laboratory Surveillance System for Streptococcus pneumoniae isolated of invasive disease from 2007 to 2012 and compares the incidence of invasive pneumococcal disease (IPD) by age groups in the prevaccinal (2007-2010) and postvaccinal period (2012). Methods: Descriptive study of S. pneumoniae surveillance in invasive diseases cases confirmed at the National Reference Laboratory of the Institute of Public Health of Chile from 2007 to 2012. Results: Global incidence of laboratory confirmed IPD cases decreased 27.8% from 2007 to 2012 and showed a lower risk for IPD in 2012 compared with 2007. Incidence in children aged 1 year or less decreased from 56.1 to 16.3 per 100,000 and from 42.0 to 19.9 per 100,000 in children aged 12 to 23 months in the same period. Highest decreases were observed in IPD cases caused by serotypes 4 (100%), 19F (93.3%), 23F (90.9%), 14 (81.1%), 6B (70%), 18C (58.3%) and 1(81.8%) in children aged 2 years or less. Conclusion: Surveillance System detects S.pneumoniae isolated from invasive diseases, contributing with information about laboratory confirmed IPD trends, prevalent serotypes and replacement effects. These results can be used as evidence in healthcare decision making for pneumococcal vaccines.

Introducción: La vacuna neumocóccica 10 valente fue incorporada al Programa Nacional de Inmunizaciones (PNI) desde enero de 2011 para lactantes mediante un esquema de cuatro dosis, y desde 2012, con un esquema de tres dosis. El objetivo de esta publicación es dar a conocer el resultado de la vigilancia de laboratorio de Streptococcus pneumoniae aislado de enfermedad invasora (ENI) desde el año 2007 al 2012 y comparar la incidencia de esta enfermedad según grupos de edades en un período prevacunal (2007-2010) con el postvacunal (2012). Materiales y Métodos: Estudio descriptivo de los resultados de la vigilancia de S. pneumoniae en los casos de ENI confirmados microbiológicamente en Chile, en el Laboratorio Biomédico Nacional de Referencia del Instituto de Salud Pública de Chile (ISP) durante los años 2007 a 2012. Resultados: La evolución de la incidencia global de S. pneumoniae en casos de ENI muestra un menor riesgo en los años estudiados (OR 2011 vs 2007-2010: 0,82 (IC 95%: 0,75-0,89); OR 2012 vs 2007-2010: 0,76 (IC 95%: 0,70-0,82)). En niños bajo un año de edad, la incidencia disminuyó desde 56,1 a 16,3 por 100.000 y en niños de 12 meses a 23 meses desde 42,0 a 19,9 por 100.000, en el mismo período. Los mayores porcentajes de disminución en los menores de 2 años se observaron en los casos de ENI producidos por los serotipos 4 (100%), 19F (93,3%), 23F (90,9%), 14 (81,1%), 6B (70%), 18C (58,3%) y 1(81,8%). Conclusión: El sistema de vigilancia permite detectar cepas de S. pneumoniae aisladas de enfermedad invasora en nuestro país, lo que aporta información respecto de la tendencia de la ENI confirmada microbiológicamente en Chile, los serotipos prevalentes y el posible efecto de reemplazo de ellos descrito en otros países, aportando a la autoridad de salud una herramienta adicional para la toma de decisiones respecto del tipo de vacuna a usar en el PNI con la mejor evidencia disponible.

Vigilancia de laboratorio de enfermedad meningocóccica invasora en Chile, 2006-2012 / Laboratory surveillance for invasive meningococcal disease in Chile, 2006-2012

Background: Laboratory surveillance of Invasive Meningococcal Disease (IMD) is performed by the Institute of Public Health of Chile. It confirms identification, classifies in serogroups and analyzes the genetic profiles of Neisseria meningitidis isolates from laboratories throughout the country. Aim: To show the results of this surveillance from 2006 to 2012. Methods: A descriptive data analysis of the confirmed cases of IMD and serological characterization, susceptibility and genetic profiles of the isolates. The analysis was disaggregated by serogroup, age and region. Results: From 2006 to 2012, 486 isolates of N. meningitidis were confirmed. In 2011 a rise in IMD rates was observed due to an increase in W serogroup cases, mainly affecting children aged 5 years or less. Serogroup W became the most prevalent during 2012 (58.3%), replacing the historically prevalent serogroup B. Predominating strains belonged to ST-32 complex/ET-5 complex (40, 4% of strains) and ST-41/44 complex/ Lineage 3 (45, 9% of strains). Conclusions: Laboratory surveillance has allowed the early detection of increasing IMD caused by serogroup W, which is emergent in Chile. This information has reinforced the daily monitoring of new cases, in collaboration with all the clinical laboratories of the country.

Introducción: La vigilancia de laboratorio de enfermedad meningocócica invasora (EMI) que realiza el Instituto de Salud Pública de Chile, confirma, seroagrupa y estudia el perfil genético de las cepas de Neisseria meningitidis provenientes de los laboratorios del país. Objetivo: En este artículo se muestra los resultados de esta vigilancia entre los años 2006 a 2012. Materiales y Métodos: Se realizó un análisis descriptivo de los casos confirmados de EMI, caracterización serológica, el análisis de susceptibilidad antimicrobiana y el estudio de subtipo genético de la cepa. El análisis se desagregó por serogrupo, edad y región. Resultados: En el período 2006-2012 fue confirmado un total de 486 cepas de N. meningitidis. A partir del año 2011 se observó un alza en la tasa de EMI dado por el número de casos del serogrupo W, afectando principalmente a niños bajo 5 años de edad. El W se transformó en el serogrupo prevalente el año 2012 (58,3%), desplazando al serogrupo B, el cual históricamente había sido prevalente. Predominaron principalmente las cepas pertenecientes al complejo clonal ST-32 complex/ET-5 complex (40,4% de las muestras) y el ST-41/44 complex/Lineage 3 (45,9% de las muestras). Conclusiones: El sistema de vigilancia de laboratorio ha permitido la identificación del serogrupo W, emergente en Chile. Esta información nos ha obligado a estar en permanente alerta y monitoreo de casos diarios, mediante la participación activa de todos los laboratorios clínicos del país.

Inicio del brote de Influenza A (H1N1) pandémica en Chile: caracterización genética de los primeros casos detectados / Genetic characterization of the virus causing H1N1 influenza pandemic in Chile: Analysis of the first detected cases

Background: Following the announcement of the Influenza A(H1N1) pandemic by the World Health Organization in April 2009, a surveillance program was carried out in Chile to detect the introduction of the virus in the country and to monitor its propagation and impact. Aim: To describe the onset of the outbreak and the genetic characterization of the pandemic H1N1 influenza virus in the first detected cases in Chile. Material and Methods: Analysis of18 clinical samples coming from suspicious patients, received in a National Reference Laboratory. RNA reverse transcription and real time influenza gene DNA amplification was carried out in a 7500 Fast and Step One Real Time PCR Systems of Applied Biosystems and MxPro-Mx3000P thermocycler from Stratagene. Super Script III Platinum One-Step Quantitative RT-PCR was used. Results: The virus was first detected in three persons returning from the Dominican Republic via Panamá and a child from the east zone of Santiago. Genetic characterization of the virus showed that the child was infected by a different variant of the pandemic virus than the three persons returning from the Caribbean. Conclusions: The onset of the Influenza outbreak in Chile apparently carne from two different epidemiological groups. The spread of the virus detected in the voyagers was limited immediately However the virus of the fourth case was found in different regions of Chile.

Detección de mutaciones asociadas a cepas multidrogo resistente de Mycobacterium tuberculosis en Chile / Detection of genes associated with drug resistance in Mycobacterium tuberculosis strains isolated in Chile

Background: The incidence of acquired resistance to antituberculous drugs of Mycobacterium tuberculosis in Chile is approximately 23 percent. Aim: To analyze the mutations associated with drug resistance in drug resistant strains of Mycobacterium tuberculosis. Material and Methods: In 28 drug resistant Mycobacterium tuberculosis strains isolated in Chile, genes leading to drug resistance were studied. DNA was amplifed by polymerase chain reaction (PCR) and sequencing was carried out using the ABI PRISM big dye terminator cycle sequencing ready reaction kit. Results: In rifampicin-resistant strains, the mutations in rpoβ gene were in the codons S531W/L (56 percent), D516Y (16 percent) and D516V (16 percent). The predominant mutation in katG gene was in the codon S315L (73 percent) in isoniazid-resistant strains. The mutation S95T was found in the 71 percent of ciprofoxacin resistant strains. Only one ethambutol resistant strain had the M306I mutation. Three unreported mutations in katG were identifed. Conclusions: Drug resistance associated mutations of Mycobacterium tuberculosis isolated in Chile were similar to those reported abroad.

Staphylococcus aureus comunitario resistente a cloxacilina: comunicación de los primeros cinco casos descritos en Chile / Community acquired infections with methicillin resistant strains of Staphylococcus aureus: report of five cases

Community acquired infections with methicillin resistant strains of Staphylococcus aureus (MRSA) infections have a more aggresive clinical course and involve mostly skin and lungs. These infections appear as outbreaks among prisoners, sportsmen, men having sex with men and military personnel. The higher aggressiveness of these strains is due to the production of several toxins, mainly Panton- Valentine leukocidine. The detection of the gene that codes for this toxin is a distinctive feature of these strains. We report five patients with community acquired MRSA infections. The clinical presentation was a skin infection in all. One patient had a pleuropneumonia in addition. Apart for resistance to beta-lactam antimicrobials, the strains were resistant to erythromycin and ciprofloxacin. Patients were treated with vancomycin, clotrimoxazole or intravenous clindamycin with a good evolution. An epidemiológical surveillance for community acquired MRSA strain infections should be started and measures to adequately treat infected patients and avoid dissemination should be implemented.

Caracterización molecular en aislados chilenos de Streptococcus agalactiae / Molecular characterization of Chilean isolates of streptococcus agalactiae

Streptococcus agalactiae is the main causing organism of invasive infections such as sepsis and meningitis in the newborn. Aim: To perform a genotype characterization of Streptococcus agalactiae strains coming form invasive infections of newborns and colonized pregnant women. Material and methods: A group of 58 strains not related epidemiologically isolated from colonized pregnant women and invasive infections in newborns, were studied. Pulsed field electrophoresis (PFGE) and polymerase chain reaction amplification of hylB and IS 1548 genes, as possible virulence markers, were performed. Results: Among the studied strains, 37 genetic subtypes were observed. There were nine groups of identical PFGE patterns. Three corresponded to serotype la and six to serotype III. An erythromycin and clindamycin resistant clone was identified in three colonized women and a newborn with sepsis, which were not epidemiologically related. The hylB gene was equally present in cases of neonatal meningitis or colonized pregnant women. Conclusions: There was a great degree of polymorphism among the studied strains. The ample presence of hylB gene and the absence of the insertion element IS1548 in the hylB gene in invasive and colonizing strains, indicates that both groups of strains are potentially pathogenic.

The bacteriophage lambda DNA packaging enzyme: identification of four structural domains of the gpNu1 subunit using limited proteolysis

Lambda DNA terminase, the enzyme that cleaves virion-length chromosomes from multigenomic concatemers and packages them into the bacteriophage head, is composed of two subunits, gpNu1 and gpA. Direct determination of the structure of gpNu1, the smaller subunit, has not been possible because of its insolubility in aqueous solutions. Therefore, to identify smaller and potentially water-soluble domains of gpNu1, we analyzed the nature of the products obtained by limited digestion of the protein with several proteases. The gpNu1 subunit was obtained from E. coli cells transfected with the plasmid pH6-Nu1 that overproduces the protein. Incubation of gpNu1 solubized in 2.5 M guanidinium chloride with chymotrypsin resulted in the formation of at least eight discrete protein bands, while treatment with endoproteinase glu-C and bromelain yielded three and one major bands, respectively. The peptides generated by digestion with the various proteases were separated by two-dimensional gel electrophoresis and transferred to Immobilon membranes. Amino acid sequencing of the peptides allowed for the precise assignment of their N-terminal amino acid, while their estimated molecular weights permitted the identification of their C-terminal ends. The results reveal that in the presence of 2.5 M guanidinium chloride, gpNu1 is partially folded in at least four distinct structural domains that correspond to functional domains as determined by previously reported genetic experiments. This information is key to design new plasmids to overproduce these domains for further structural analysis.