Diego Vicente

Viruses in community-acquired pneumonia in children aged less than 3 years old: High rate of viral coinfection.

The occurrence of viral coinfections in childhood pneumonia has received little attention, probably because suitable detection methods have been lacking. Between November 2004 and October 2006, the presence of 14 respiratory viruses in children aged less than 3 years old with community‐acquired pneumonia were investigated using molecular or immunochromatographic techniques and/or viral culture. A total of 315 children (338 episodes) were included, and hospitalization was required in 178 episodes. At least one virus was detected in 66.9% of the episodes and simultaneous detection of two or more viruses was frequent (27% of the episodes with viral detection). The most frequently detected virus was respiratory syncytial virus (n = 67: 33 subgroup A, 33 subgroup B, 1 not typed), followed by human bocavirus (n = 48), rhinovirus (n = 46), human metapneumovirus (n = 39: 13 genotype A2, 8 B1, 5 B2, 1 A1, 12 not genotyped) and parainfluenza viruses (n = 38: 1 type 1, 3 type 2, 22 type 3, 11 type 4 and 1 not typed). The 14 viruses investigated were found in viral coinfections, which were more frequent in children aged less than 12 months. Except for adenovirus, the incidence of which was low, the percentage of viral coinfection ranged between 28.2% and 68.8%. Children with viral coinfection more frequently required hospital admission than those with single viral infection. It is concluded that viral coinfections are frequent in children aged less than 3 years old with community‐acquired pneumonia and can be a poor prognostic factor. J. Med. Virol. 80:1843–1849, 2008.

Human Bocavirus, a Respiratory and Enteric Virus

In Spain, human bocavirus (HBoV) was detected in 48 (9.1%) of 527 children with gastroenteritis at similar frequency as for children with respiratory illness (40/520, 7.7%). Fecal excretion adds new concern about the transmission of HBoV. To our knowledge, this report is the first to document HBoV in human feces.

Human parechoviruses in infants with systemic infection

Human parechoviruses (HPeVs) are RNA viruses related to neonatal sepsis, meningoencephalitis and other infections in young children. Little clinical and epidemiological information is available on these viruses. HPeVs were sought in cerebrospinal fluid from 397 infants aged less than 12 months from whom a sample was obtained to exclude meningitis or encephalitis from 2006 to 2009. HPeV infections were also tested in stool samples from 271 children aged less than 3 years old with gastroenteritis from November 2008 to March 2009. HPeV detection was by real‐time polymerase chain reaction assay (region 5′UTR), followed by genotyping (region VP3/VP1). HPeVs were detected in the cerebrospinal fluid of nine infants (2.3%), one aged 6 months and eight aged 14–55 days old. All were admitted to hospital for febrile syndrome with abrupt clinical deterioration and suspected systemic infection without clear laboratory signs of meningeal inflammation. The same virus was detected in all the available nasopharyngeal aspirates, stool, and/or serum samples from each patient. At least eight of the nine cases were caused by HPeV3. HPeVs were detected in stool samples from 17 children (6.3%), the most prevalent types being types 1 and 3. In conclusion, HPeV infection is common in the Basque Country (Spain) and HPeV3 is a significant cause of hospital admission due to systemic infection in the first few months of life. In these patients, HPeVs should be investigated as part of routine tests for enterovirus. J. Med. Virol. 82:1790–1796, 2010.

Human Metapneumovirus and Chronic Obstructive Pulmonary Disease

To the Editor: We read with interest an article, Human Metapneumovirus Detection in Patients with Severe Acute Respiratory Syndrome, in your journal (1). In the report, Chan et al., did not question that SARS-CoV is the etiologic agent of severe acute respiratory syndrome (SARS); however, human metapneumovirus (HMPV) was found in 25 (52%) of 48 probable SARS cases that were investigated, and SARS-CoV was detected in 11 (22.9%) of them. Another recent article reported HMPV in five of six patients in whom SARS was diagnosed in Canada (2); four of the six were coinfected with SARS-CoV. The prevalence of HMPV infection in SARS patients validates the interest in HMPV’s possible role in SARS etiology. From November 2001 to February 2002, 1 year before the first cases of SARS appeared, we tested the sputum of patients >64 years of age who had experienced exacerbation of chronic obstructive pulmonary disease, for HMPV. Investigations were conducted on 90 episodes in 89 elderly patients, 62 males and 27 females, in which we found no other microorganisms that could have been related to the exacerbation of chronic obstructive pulmonary disease. RNA was extracted from the sputum samples and amplified by reverse transcriptase–polymerase chain reaction (RT-PCR) to detect HMPV as previously described (3). Results of bacterial culture and culture and PCR to detect respiratory syncytial virus and influenza virus types A and B were negative, whereas HMPV was found in the sputum of five (three men and two women) immunocompetent patients, 77–87 years of age. The prevalence of HMPV infection was 5.5%, similar to the percentage obtained by Chan et al., when HMPV RT-PCR was conducted on the respiratory samples. Fever (temperature >38°C) was not present in any of the five patients infected with HMPV. Two patients were admitted to a hospital. Both patients had bronchial infection and cough with bronchospasm and moderate respiratory insufficiency (oxygen saturation rate: 90.3% and 88%, respectively) for >1 week. Sputum samples from an additional 70 elderly patients with exacerbation of chronic obstructive pulmonary disease with positive detection for influenza virus (n = 50) or respiratory syncytial virus (n = 20) were tested for HMPV infection. None of the samples showed HMPV infection. Sequence analysis of amplicons from the five samples positive for HMPV infection showed >95% similarity with HMPV sequences found in other parts of the world (4,5). Additional studies should be conducted to confirm that HMPV exacerbates chronic obstructive pulmonary disease. However, by performing an RT-PCR directly on the sample instead of the more efficient RT-PCR after viral culture used by Chan et al., these findings suggest that HMPV is a frequently undetected agent in acute respiratory infection unrelated to SARS. The important questions are whether HMPV and SARS-CoV coinfection would facilitate more severe SARS, or whether HMPV infection would facilitate a more efficient transmission of SARS-CoV.

Características clínicas de los niños hospitalizados por infección por virus Influenza

Antecedentes Las manifestaciones clinicas de la gripe son bastante inespecificas y similares a otras infecciones virales, a procesos respiratorios de otra etiologia e incluso a cuadros septicos en lactantes. Existen pocos estudios sobre las caracteristicas clinicas de la infeccion por virus Influenza en ninos hospitalizados. Objetivo Evaluar las caracteristicas clinicas de los ninos hospitalizados por infeccion por virus Influenza en cuatro ondas epidemicas consecutivas (2000-2004). Material y metodos Estudio retrospectivo por revision de historias clinicas de los ninos hospitalizados por infeccion confirmada por virus Influenza A y B, mediante cultivo celular y reaccion en cadena de la polimerasa. Se registraron las variables:sexo, edad, clinica al ingreso, exploraciones complementarias,diagnostico y evolucion posterior. Resultados Fueron hospitalizados 84 ninos, 74 casos debidos a infeccion por virus Influenza AH3, 5 casos por virus Influenza AH1 y cinco por Influenza B. El 42,8% fueron menores de 6 meses. El cuadro clinico principal fue fiebre (75 casos), tos (56 casos), afectacion otorrinolaringologica (53 casos). Los diagnosticos mas frecuentes que motivaron el ingreso fueron sindrome febril (75 casos), bronquiolitis (19 casos), neumonia (13 casos) y bronquitis (8 casos). En 21 casos se objetivo coinfeccion viral o bacteriana,siendo la asociacion con virus respiratorio sincitial la coinfeccion mas frecuente (10 casos). Se han observado pocas diferencias entre los diferentes grupos de edad, excepto para neumonia y fiebre prolongada (mas frecuentes en mayores de 6 meses) y linfocitosis (en menores de 6 meses). El cuadro febril agudo en menores de 6 meses ha originado mayor numero de exploraciones complementarias en los de menor edad. Solo en tres ninos se encontraron factores de riesgo para hospitalizacion,todos ellos mayores de 6 meses. Todos los casos evolucionaron favorablemente. Conclusiones La infeccion por virus Influenza en los ninos que estan hospitalizados predomina en lactantes sanos, con fiebre y signos y sintomas respiratorios poco especificos, similares a otros cuadros infecciosos. Es necesario aplicar tecnicas diagnosticas microbiologicas especificas para el diagnostico precoz. Los lactantes sanos y menores de 24 meses constituyen el grupo de mayor riesgo para la hospitalizacion por infeccion por virus Influenza.

Molecular Characterization of Erythromycin-Resistant Clinical Isolates of the Four Major Antimicrobial-Resistant Spanish Clones of Streptococcus pneumoniae (Spain23F-1, Spain6B-2, Spain9V-3, and Spain14-5)

Erythromycin resistance and the characterization of the corresponding determinants of resistance were studied in clinical Streptococcus pneumoniae isolates belonging to the four major multiresistant pneumococcal Spanish clones (ermB and mefA genes for the Spain23F-1, Spain9V-3, serotype 14 variant of the Spain9V-3 and Spain14-5 clones and ermB gene for the Spain6B-2 clone). These isolates were confirmed as major clones by pulsed-field gel electrophoresis (PFGE), BOX-PCR, and multilocus sequence typing (MLST). The spread and prevalence of these erythromycin-resistant variants of the Spanish clones in an area of the north of Spain were dissimilar-low for the Spain9V-3 clone (5.8% among the isolates belonging to this clone, including isolates of the serotype 14 variant) and very frequent for the Spain14-5 clone (91.7%).

High Rate of Fecal Carriage of Extended-Spectrum-β-Lactamase-Producing Escherichia coli in Healthy Children in Gipuzkoa, Northern Spain

ABSTRACT The prevalence of extended-spectrum-β-lactamase-producing Enterobacteriaceae (ESBLPE) was studied in stool samples from 125 8- to 16-month-old healthy children. Twenty-four percent of them and 10.7% of the 318 fecal samples studied yielded extended-spectrum-β-lactamase-producing Escherichia coli, with the types being SHV-12, CTX-M-1, CTX-M-14, and TEM-52, the most common types of β-lactamases. This high prevalence of ESBLPE in healthy people, which is to our knowledge the highest currently reported in Europe, may represent a risk for increased infections by these organisms in the future.

Fatal Neisseria meningitidis serogroup X sepsis in immunocompromised patients in Spain. Virulence of clinical isolates

BACKGROUND Neisseria meningitidis X (NmX) is a minority meningococcus serogroup for which no vaccine is available. Huge meningococcus X outbreaks occurred in West Africa but developed countries have reported only sporadic cases. Two invasive fatal cases are presented. METHODS Isolates were characterized by serogrouping (latex agglutination and genogroup X-specific polymerase-chain-reaction), PorA and FetA typing, multilocus sequence typing and pulsed-field gel electrophoresis. Immunocompetent female balm/C mice inoculated intraperitoneally were use to test virulence of invasive and carrier isolates. RESULTS Until April 2010, NmX was absent among 868 invasive meningococci characterized in the Basque Country in a 20-year period. In April 2010, two fatal NmX: P1.21,16: F5-5/ST750 episodes were detected in unrelated immunodeficient patients. After analysis of 803 meningococcal isolates from asymptomatic carriers obtained between 1988 and 2010, eight NmX isolates were detected. The genotype of the two invasive isolates bore no relation to any of the NmX isolates detected in healthy individuals from the Basque Country or to isolates from outbreaks in Africa. CONCLUSIONS NmX isolates in the north of Spain can cause severe disease in humans, despite their low prevalence. The in vivo animal study showed that virulence of isolates was more closely associated with the genotype than with the serogroup.

Detection of bacteria and viruses in the pleural effusion of children and adults with community-acquired pneumonia

AIM To study the etiology and the utility of new molecular methods in the diagnosis of complicated pneumonia with empyema. MATERIALS & METHODS Bacteria and viruses detection was performed by several traditional and molecular methods in the pleural fluid (PF) of 60 patients (38 children) with community-acquired pneumonia (CAP). RESULTS Despite prior antimicrobial therapy in 49 (81.7%) CAP patients, an etiological diagnosis could be established in 41 (68.3%), 35 being (58.3%) Streptococcus pneumoniae. PF culture was positive in only 6 patients but each molecular test detected more than 82% of cases. CONCLUSION Traditional culture methods have poor diagnostic sensitivity in PF because most CAP patients are under antimicrobial therapy when it is obtained. S. pneumoniae detection by molecular methods highly improves diagnosis.

Oseltamivir-resistant pandemic influenza a (H1N1) 2009 viruses in Spain

BACKGROUND Pandemic influenza A (H1N1) 2009 virus appeared in Spain on April 25, 2009 for the first time. This new virus was adamantane-resistant but it was sensitive to neuraminidase (NA) inhibitors oseltamivir and zanamivir. OBJECTIVES To detect oseltamivir-resistant pandemic influenza A (H1N1) 2009 viruses by the Spanish Influenza Surveillance System (SISS) and a possible spread of oseltamivir-resistant viruses in Spain since starting of the pandemic situation. STUDY DESIGN A total of 1229 respiratory samples taken from 413 severe and 766 non-severe patients with confirmed viral detection of pandemic influenza A (H1N1) 2009 viruses from different Spanish regions were analyzed for the specific detection of the H275Y mutation in NA between April 2009 and May 2010. RESULTS H275Y NA substitution was found in 8 patients infected with pandemic influenza A (H1N1) 2009 viruses collected in November and December 2009 and in January 2010. All oseltamivir-resistant viruses were detected in severe patients (8/413, 1.93%) who previously received treatment with oseltamivir. Six of these patients were immunocompromised. CONCLUSION In Spain, the number of oseltamivir-resistant pandemic influenza A (H1N1) 2009 viruses is until now very low. No evidence for any spread of oseltamivir-resistant H1N1 viruses is achieved in our Country.