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.

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.

Bacteremic pneumococcal pneumonia: serotype distribution, antimicrobial susceptibility, severity scores, risk factors, and mortality in a single center in Chile

AIMS: Bacteremic pneumococcal pneumonia (BPP) is a severe condition. To evaluate seasonal distribution, mortality, serotype frequencies, antimicrobial susceptibility, and different severity scores among patients with BPP. PATIENTS AND METHODS: Patients were identified by laboratory data and restricted to adulthood. Standard methods were used for serotyping and antimicrobial susceptibility. Risk factors were analyzed by univariate and multivariate methods. Severity scores (APACHE II, CURB-65 and CAP PIRO) were compared using ROC curves. RESULTS: Sixty events of community-acquired BPP occurred between 2005 and 2010. A seasonal pattern was detected. Mean age was 72.1 years old (81.4% >60 years). All had a predisposing factor. Previous influenza (3.3%) or pneumococcal immunization (1.7%) was infrequent. Admission to critical units was required by 51.7%. Twenty-two serotypes were identified among 59 strains. Only one strain had intermediate resistance to penicillin (1.7%). In-hospital mortality reached 33.3%. Multivariate analysis identified a CAP PIRO score>3 (OR 29.7; IC95 4.7-187), age >65 years (OR 42.1; IC95 2.2-796), and a platelet count<100,000/μL (OR 10.9; IC95 1.2-96) as significant independent factors associated with death. ROC curve analysis did not reveal statistical differences between the three severity scores to predict death (AUC 0.77-0.90). The prognostic yield for all of them was limited (Positive Likelihood Ratio: 1.5-3.8). CONCLUSIONS: BPP had a high case-fatality rate in this group of adult patients with no association to resistant isolates, and a low immunization record. Three independent factors were related to death and the prognostic yield of different severity scores was low. .

Subtipificación molecular de Salmonella entérica serotipo Enteritidis en el período post epidémico / Molecular subtyping of Salmonella enterica serotype Enteritidis in a post epidemic period

Background: In the last two decades, Salmonella enterica serotype Enteritidis has become one of the main agents causing food borne diseases worldwide. This agent is transmitted mainly by contaminated meat and poultry. Aim: To determine the genetic subtypes of Salmonella enterica serotype Enteritidis, circulating in Chile between 2001 and 2003, a post epidemc period. Material and methods: One hundred ninety three isolates coming from human samples, prepared foods and animal products for human consumption, were analyzed bypulsed field electrophoresis, using PulseNet standardized protocol. Results: Thirteen subtypes of Salmonella enterica serotype Enteritidis were identified, that had between 0 and 13 bands. A predominant subtype was identified in 172 strains (88%) that carne from human isolates, prepared foods and animal producís for human consumption. Other four subtypes, found in prepared foods and animal products for human consumption, were also found in human isolales. Most subtypes were lighlly inlerrelaled Subtypes II, VIII and XI were also found in the 1994 epidemic. Conclusions: Subtyping of baclerial strains by pulsed field electrophoresis is useful for the surveillance of food borne diseases.

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.