Full-length genome characterization and genetic relatedness analysis of hepatitis A virus outbreak strains associated with acute liver failure among children.

Clinical infection by hepatitis A virus (HAV) is generally self-limited but in some cases can progress to liver failure. Here, an HAV outbreak investigation among children with acute liver failure in a highly endemic country is presented. In addition, a sensitive method for HAV whole genome amplification and sequencing suitable for analysis of clinical samples is described. In this setting, two fatal cases attributed to acute liver failure and two asymptomatic cases living in the same household were identified. In a second household, one HAV case was observed with jaundice which resolved spontaneously. Partial molecular characterization showed that both households were infected by HAV subtype IA; however, the infecting strains in the two households were different. The HAV outbreak strains recovered from all cases grouped together within cluster IA1, which contains closely related HAV strains from the United States commonly associated with international travelers. Full-genome HAV sequences obtained from the household with the acute liver failure cases were related (genetic distances ranging from 0.01% to 0.04%), indicating a common-source infection. Interestingly, the strain recovered from the asymptomatic household contact was nearly identical to the strain causing acute liver failure. The whole genome sequence from the case in the second household was distinctly different from the strains associated with acute liver failure. Thus, infection with almost identical HAV strains resulted in drastically different clinical outcomes.

Multidrug-resistant tuberculosis among patients in Baja California, Mexico, and Hispanic patients in California.

OBJECTIVES: We sought to compare prevalence and determinants of multidrug-resistant tuberculosis (MDR-TB) between tuberculosis patients in Baja California, Mexico, and Hispanic patients in California. METHODS: Using data from Mexico's National TB Drug Resistance Survey (2008-2009) and California Department of Public Health TB case registry (2004-2009), we assessed differences in MDR-TB prevalence comparing (1) Mexicans in Baja California, (2) Mexico-born Hispanics in California, (3) US-born Hispanics in California, and (4) California Hispanics born elsewhere. RESULTS: MDR-TB prevalence was 2.1% in Baja California patients, 1.6% in Mexico-born California patients, 0.4% in US-born California patients, and 2.7% in Hispanic California patients born elsewhere. In multivariate analysis, previous antituberculosis treatment was associated with MDR-TB (odds ratio [OR] = 6.57; 95% confidence interval [CI] = 3.34, 12.96); Mexico-born TB patients in California (OR = 5.08; 95% CI = 1.19, 21.75) and those born elsewhere (OR = 7.69; 95% CI = 1.71, 34.67) had greater odds of MDR-TB compared with US-born patients (reference category). CONCLUSIONS: Hispanic patients born outside the US or Mexico were more likely to have MDR-TB than were those born within these countries. Possible explanations include different levels of exposure to resistant strains and inadequate treatment.

Evolution of dengue virus in Mexico is characterized by frequent lineage replacement.

Both dengue fever and its more serious clinical manifestation, dengue hemorrhagic fever, represent major public health concerns in the Americas. To understand the patterns and dynamics of virus transmission in Mexico, a country characterized by a marked increase in dengue incidence in recent years, we undertook a molecular evolutionary analysis of the largest sample of Mexican strains of dengue virus compiled to date. Our E gene data set comprises sequences sampled over a period of 27 years and representing all of the Mexican states that are endemic for dengue. Our phylogenetic analysis reveals that, for each of the four dengue viruses (DENV-1 to DENV-4), there have been multiple introductions of viral lineages in Mexico, with viruses similar to those observed throughout the Americas, but there has been strikingly little co-circulation. Rather, dengue virus evolution in Mexico is typified by frequent lineage replacement, such that only a single viral lineage dominates in a specific serotype at a specific time point. Most lineage replacement events involve members of the same viral genotype, although a replacement event involving different genotypes was observed with DENV-2, and viral lineages that are new to Mexico are described for DENV-1, DENV-3 and DENV-4.

[Analysis of nosocomial pediatric bacteremias at a general hospital between 1990 and 2006. The impact of attending the intravascular therapy]. / Análisis de las bacteriemias nosocomiales pediátricas en un hospital general entre 1990 y 2006. Impacto de la atención a la terapia intravascular.

INTRODUCTION: The administration of parenteral infusates is a frequent intervention that is considered innocuous; moreover, the risk of this procedure which offers a direct access to the bloodstream is minimized. OBJECTIVE: To evaluate the epidemiology of nosocomial pediatric bacteremias after implementing a control program. METHODS: Analysis of pediatric bacteremias was made in 3 periods: 1) 1990-1992, prior to establishing strategies to avoid contamination of parenteral infusions; 2) 1996, the phase after establishing these strategies; and 3) 2005-2006, the recent situation in the hospital. RESULTS: The proportion of gram-negative rods isolated in blood cultures dropped from 82.9 to 35.1% (p = 0.004) during the 17-year study period. There was no significant difference in the proportion of gram-negative rods isolated from intravascular catheters. The proportion of contaminated parenteral infusions dropped from 22.2% to 0.4% (p < 0.001). DISCUSSION: The strategies established to avoid the contamination of parenteral infusions were associated with a reduction in the proportion of gram-negative rods in blood cultures, although the proportion is still higher than that in developed countries, probably related to catheter contamination. We suggest establishing similar strategies in other hospitals from developing countries.

Global and local variations in antimicrobial susceptibilities and resistance development in the major respiratory pathogens.

Acute respiratory tract infections, such as bacterial pneumonia and acute exacerbations of chronic bronchitis, have been identified by the World Health Organisation as the leading global infectious cause of death. An increasing prevalence of antibiotic resistance has been identified worldwide in the three major bacterial respiratory pathogens -Streptococcus pneumoniae, Moraxella catarrhalis and Haemophilus influenzae. The selection and spread of resistance is to some degree inevitable and the importance of monitoring its progress has led to the instigation of numerous international, regional and national surveillance programmes. The results from surveillance studies show wide variations in susceptibility rates, both geographically and over time, highlighting the need for local resistance prevalence data in order to guide empirical prescribing and to identify areas in which medical need for new agents is greatest.

[Molecular characterization of an outbreak caused by CTX-M-12-producing Klebsiella pneumoniae in a Colombian hospital's neonatal intensive care unit]. / Caracterización molecular de un brote por Klebsiella pneumoniae productora de CTX-M-12 en la unidad de cuidado intensivo neonatal de un hospital Colombiano.

INTRODUCTION: Molecular characterisation of Klebsiella pneumoniae strains is a tool that assits in the reduction of the disemination of drug resistance and the control of nosocomial infections that are caused by this pathogen. Objective. Molecular description of an outbreak of nosocomial infection caused by Klebsiella pneumoniae in a neonatal intensive care unit in a tertiary level hospital in Bogotá. METHODS: Eleven Klebsiella pneumoniae isolates were analysed. Production of Extended Spectrum Beta-Lactamases was verified by agar diffusion tests. Isoelectric points of the enzymes were determined by isoelectric focusing. The bla(CTX-M-12) gene was detected by PCR and pulsed field gel electrophoresis genotyping was done. RESULTS: All the isolates were Extended Spectrum Beta-Lactamase producers. Pulsed field gel electrophoresis and BOX-PCR genotyping grouped two isolates from hospital objects and eight infection-causing isolates into a single epidemic clone. The isolate from a thermometer was not grouped into the epidemic clone and showed a different resistance pattern. Isoelectric focusing revealed simultaneous beta-lactamase production having different isoelectric points. PCR amplification revealed the presence of the bla(CTX-M-12) gene in the 11 isolates studied. CONCLUSION: This is the first report of a molecularly characterised outbreak of CTX-M-12-producing Klebsiella pneumoniae from Colombia. The results of this study provide additional evidence of the global dissemination of CTX-M ESBL and the need for epidemiological follow-up in our hospitals.

Nosocomial bacteremia in neonates related to poor standards of care.

BACKGROUND: In developing countries, intravenous liquids are mixed and administered by nurses, sometimes under suboptimal infection control conditions. We hypothesized that outbreaks of infusate-associated neonatal bacteremias are common, and we evaluated whether they can be detected by vigilant microbiologic surveillance of infusates. METHODS: We studied intravenous infusates administered to neonates in a Mexican hospital where mixtures of infusates were prepared in hospital wards. The study was performed in 3 stages: stage 1, initial culturing of in-use infusates under basal conditions; stage 2, prospective culturing during a cluster of clinical sepsis; and stage 3, final culturing once the outbreak was controlled. RESULTS: In stage 1, 68 infusates were sterile, and 1 was contaminated with Staphylococcus aureus (1.45%), from 23 patients. In stage 2, of 182 infusates from 39 patients, 51 infusates (28%) were contaminated with Gram-negative rods. On the first day of stage 2, 11 of 15 infusates were contaminated with the same strain of Klebsiella pneumoniae, which continued to appear for 26 days. Another 4 strains of Gram-negative rods were also isolated during stage 2. The association between contaminated infusate and death was significant (odds ratio, 9.4; 95% confidence interval, 2-44.3; P < 0.001). Mixtures made by nurses were more likely contaminated than commercial preparations (odds ratio, 3.1; 95% confidence interval, 1.1-8.5; P = 0.037). In stage 3, there were 42 sterile infusates from 22 patients. CONCLUSIONS: Our study suggests that poor standards of care common in hospitals from developing countries sometimes result in outbreaks of sepsis and death for newborn patients.

[Molecular epidemiology of nosocomial infection by extended-spectrum beta-lactamases-producing Klebsiella pneumoniae]. / Epidemiología molecular de infección nosocomial por Klebsiella pneumoniae productora de beta-lactamasas de espectro extendido.

Molecular epidemiology applied to the study of nosocomial infection has been fundamental in formulating and evaluating control methods. From patients in a level 3 Bogota hospital, Klebsiella pneumoniae samples were isolated that produced extended-spectrum beta-lactamases (ESBL). Each of 15 isolates was characterized microbiologically and by molecular characters realized by pulsed field gel electrophoresis (PFGE) and by repetitive-DNA sequences amplification (REP-PCR). Antimicrobial susceptibility and ESBL production was determined in accordance with NCCLS guidelines. The beta-lactamases were evaluated by isoelectric-focusing and PCR. Twelve (80%) of the isolates were associated with nosocomial infection; 11 of them were from intensive care units. The antibiotic susceptibility displayed 13 resistance patterns--87% presented co-resistance to amikacin, 53% to gentamicin, 33% to ciprofloxacin, 40% to cefepime, 67% to piperacillin/tazobactam, 60% to trimethoprim/sulfamethoxazole and 47% to chloranphenicol. All were sensitive to imipenem. Production of TEM and SHV beta-lactamases was detected simultaneously in most isolates by isoelectric focusing and 93.3% produced a ceftazidimase of pl 8.2 of the SHV-5 type. The 15 isolates were grouped into 11 and 12 electrophoretic patterns by PFGE and REP-PCR, respectively. The degree of genetic variability indicated an endogenous origin of the nosocomial infections.

Caracterización molecular de un brote por Klebsiella pneumoniae productora de CTX-M-12 en la unidad de cuidado intensivo neonatal de un hospital colombiano / Molecular characterization of an outbreak caused by CTX-M-12-producing Klebsiella pneumoniae in a Colombian hospital´s neonatal intensive care unit

Introducción. La caracterización molecular de cepas de Klebsiella pneumoniae es una herramienta que contribuye a disminuir la diseminación de la resistencia y al control de las infecciones nosocomiales causadas por este patógeno. Objetivo. Describir molecularmente un brote de infección nosocomial por Klebsiella pneumoniae en la Unidad de Cuidado Intensivo Neonatal de un hospital de tercer nivel de Bogotá. Materiales y métodos. Se analizaron once aislamientos. Se verificó la producción de betalactamasas de espectro extendido mediante pruebas de difusión en agar. Se determinaron los puntos isoeléctricos de las betalactamasas mediante isoelectroenfoque. Se detectó el gen blaCTX-M-12 por PCR y se realizó genotipificación mediante BOX- PCR y electroforesis en gel con campos pulsados (PFGE). Resultados. Los aislamientos fueron productores de beta-lactamasas de espectro extendido. La genotipificación por PFGE y por BOX-PCR, agrupó a dos aislamientos provenientes de objetos hospitalarios y a los ocho aislamientos causantes de infección en un grupo clonal epidémico. El aislamiento proveniente de un termómetro no fue agrupado en el grupo clonal epidémico y mostró un patrón de resistencia diferente. Se observó la producción simultánea de beta-lactamasas con diferentes puntos isoeléctricos. La PCR reveló el gen blaCTX-M-12 en los 11 aislamientos estudiados. Conclusión: Este es el primer informe en Colombia de un brote por Klebsiella pneumoniae productora de CTX-M-12, caracterizado molecularmente. Este estudio da evidencia adicional de la diseminación global de BLEE de tipo CTX-M y alerta sobre la necesidad de actividades especificas de prevención para cortar la cadena de transmisión y del seguimiento de tipo epidemiológico en nuestros centros hospitalarios

Introduction. Molecular characterisation of Klebsiella pneumoniae strains is a tool that assits in the reduction of the disemination of drug resistance and the control of nosocomial infections that are caused by this pathogen. Objective. Molecular description of an outbreak of nosocomial infection caused by Klebsiella pneumoniae in a neonatal intensive care unit in a tertiary level hospital in Bogotá. Methods: Eleven Klebsiella pneumoniae isolates were analysed. Production of Extended Spectrum Beta-Lactamases was verified by agar diffusion tests. Isoelectric points of the enzymes were determined by isoelectric focusing. The blaCTX-M-12 gene was detected by PCR and pulsed field gel electrophoresis genotyping was done. Results. All the isolates were Extended Spectrum Beta-Lactamase producers. Pulsed field gel electrophoresis and BOX-PCR genotyping grouped two isolates from hospital objects and eight infection-causing isolates into a single epidemic clone. The isolate from a thermometer was not grouped into the epidemic clone and showed a different resistance pattern. Isoelectric focusing revealed simultaneous beta-lactamase production having different isoelectric points. PCR amplification revealed the presence of the blaCTX-M-12 gene in the 11 isolates studied. Conclusion. This is the first report of a molecularly characterised outbreak of CTX-M-12-producing Klebsiella pneumoniae from Colombia. The results of this study provide additional evidence of the global dissemination of CTX-M ESBL and the need for epidemiological follow-up in our hospitals.

Epidemiología molecular de infección nosocomial por Klebsiella pneumoniae productora de beta-lactamasas de espectro extendido / Molecular epidemiology of nosocomial infection by extended-spectrum beta-lactamases-producing Klebsiella pneumoniae

La epidemiología molecular aplicada al estudio de las infecciones nosocomiales ha sido fundamental para la formulación y la evaluación de las medidas de control; con este fin, se caracterizaron microbiológica y molecularmente aislamientos de Klebsiella pneumoniae productores de beta-lactamasas de espectro extendido (BLEE) obtenidos de pacientes en un hospital de tercer nivel de Bogotá, D.C., Colombia. Se tipificaron quince aislamientos por electroforesis en gel de campo pulsado (PFGE) y por amplificación de secuencias de AND repetidas (REP-PCR). La susceptibilidad antimicrobiana y la producción de BLEE se determinaron de acuerdo con las normas de NCCLS. Las beta-lactamasas se evaluaron por isoelectroenfoque y PCR. El 80 por ciento de estos aislamientos se asociaron con infección nosocomial y de éstos, el 91,7 por ciento provenía de unidades de cuidado intensivo. La susceptibilidad antibiótica mostró 13 patrones de resistencia; 87 por ciento de los aislamientos presentó corresistencia a amikacina, 53 por ciento a gentamicina, 33,3 por ciento a ciprofloxacina, 40 por ciento a cefepime, 66,7 por ciento a piperacilina/tazobactam, 60 por ciento trimetoprim/sulfametoxazol y 46,7por ciento a cloranfenicol. Todos fueron sensibles a imipenem. En la mayoría de los aislamientos se detectó producción simultánea de beta-lactamasas del tipo TEM y SHV y el 93,3 por ciento produjo ceftazidimasa de pI 8.2 del tipo SHV-5. Los 15 aislamientos fueron agrupados por PFGE y REP-PCR en 11 y 12 patrones electroforéticos, respectivamente. Esta variabilidad genética está relacionada con infecciones nosocomiales de origen endógeno más que por infecciones cruzadas