Milagrosa Montes

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.

Phenotypic and Genotypic Characterization of Streptococcus pyogenes Isolates Displaying the MLSB Phenotype of Macrolide Resistance in Spain, 1999 to 2005

ABSTRACT The aim of this study was to describe the genetic characteristics of Streptococcus pyogenes showing the MLSB phenotype of macrolide resistance from 1999 to 2005 in Spain and to highlight the substantial increase in these isolates in the last few years. The antimicrobial susceptibilities of 17,232 group A streptococci isolated from Madrid and Gipuzkoa from 1999 to 2005 were studied. The presence of the resistance genes ermA, ermB, mef, tetM, and tetO and the presence of the intTn and xis genes of the Tn916-Tn1545 transposon family were studied in a sample of 739 MLSB-resistant isolates. The epidemiological relationships among these isolates were analyzed by emm typing, T typing, and multilocus sequence typing. Erythromycin resistance was found in 21.3% of the isolates analyzed (annual variation of 14.3% to 28.9%). Until 2003, most erythromycin-resistant isolates showed the M phenotype, but in 2004 and 2005, about 50% of isolates showed the MLSB phenotype. Among the MLSB-resistant isolates studied, 16 clones were identified. The most prevalent clone was a strange emm11/T11/ST403 clone with a null yqiL allele. All but one of the 463 emm11/T11/ST403 isolates carried the ermB, tetM, intTn, and xis genes. The second most prevalent MLSB-resistant clone was emm28/T28/ST52, which comprised two subclones: one bacitracin-resistant, tetracycline-susceptible subclone carrying the ermB gene (n = 115) and another bacitracin-susceptible, tetracycline-resistant subclone carrying the ermB and tetM genes (n = 33). The rapid diffusion of these two clones, and especially of emm11/T11/ST403, caused the large increase in MLSB-resistant S. pyogenes isolates in Spain, suggesting a potential ability for international dissemination.

Non-endoscopic method to obtain Helicobacter pylori for culture

A sampling method to collect samples for Helicobacter pylori culture that is easier than endoscopy would be useful. We used a capsule attached to a highly absorbent nylon string, which is swallowed to obtain gastric secretions. Antral biopsy and string specimens, sequentially obtained from 36 adults with clinically suspected ulcer or gastric disease were cultured. 9 (25%) patients were negative in both tests; 18 (50%) were positive by both methods; 6 (17%) were biopsy positive and string-test negative; and 3 (8%) were biopsy negative and string-test positive. The new test is useful, simple, and causes little discomfort.

Erythromycin resistance of Streptococcus pyogenes in Madrid

BACKGROUND Erythromycin is considered to be an adequate alternative to penicillin for patients who are allergic to penicillin. Erythromycin-resistant Streptococcus pyogenes strains have been reported in some parts of the world. METHOD The in vitro activity of erythromycin and other antimicrobial agents was determined in a total of 1310 clinical Streptococcus pyogenes isolates collected in the city of Madrid from January, 1993, through December, 1996. RESULTS All strains showed susceptibility to penicillin, rifampin, vancomycin and chloramphenicol. Tetracycline resistance was 8.5%. In 36 of the strains (2.7%) MIC was 4 microg/ml for ofloxacin. Clindamycin resistance was observed in only 18 strains (1.4%); this resistance was constitutive in 15 and inducible in 3 strains. Resistance to erythromycin was observed in 14.3% of the strains, showing an increase during the study period (2.0% in 1993 vs. 22.4% in 1996; chi square for linear trend 68.8, P < 0,0001); >90% of them showed the novel resistance phenotype described by Seppälä et al. and 32 of 32 of these strains showed by PCR the 1.4-kb fragment of the mefA gene recently described as the novel macrolide efflux resistance determinant. The erythromycin-resistant strains were isolated more often in pediatric patients (144 of 872) than in adults (44 of 438) (chi square 9.9, P = 0.0016). CONCLUSION The study emphasizes the need to screen for resistance to macrolides in S. pyogenes and indicates that resistance to erythromycin in S. pyogenes has increased significantly in Madrid.

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.

Antimicrobial Susceptibility-Guided Therapy Versus Empirical Concomitant Therapy for Eradication of Helicobacter pylori in a Region with High Rate of Clarithromycin Resistance

Nonbismuth quadruple (concomitant) regimen is recommended for first‐line empirical Helicobacter pylori (HP) eradication treatment when clarithromycin resistance is more than 15–20%. Our objective was to evaluate the efficacy and tolerability of concomitant versus antimicrobial susceptibility‐guided treatment in an area with high rates of clarithromycin resistance.

Prevalence and Clonal Characterization of Streptococcus pyogenes Clinical Isolates with Reduced Fluoroquinolone Susceptibility in Spain

ABSTRACT The aim of this study was to determine the prevalence and characteristics of non-fluoroquinolone (FQ)-susceptible Streptococcus pyogenes isolates and to study their mechanisms of resistance. We performed a prospective prevalence study with 468 isolates collected from 2005 to 2007 and a retrospective study that was based on the examination of existing data collected from 1999 to 2008. The retrospective study included data for isolates with high-level resistance (HR) to ciprofloxacin (MIC ≥ 32 μg/ml) (HR isolates) and isolates with the same emm types as those reported in the literature with low-level resistance (LR) to ciprofloxacin (MICs, 2 to 8 μg/ml) (LR isolates, n = 205). Genetic characterization of the isolates was performed by means of emm typing and multilocus sequence typing. The prevalence of LR ranged from 1.9% in 2005 to 30.8% in 2007. This increase was mainly due to the circulation of an emm6 subtype (emm6.4) that represented 77.1% of the LR isolates in 2007. Notably, another emm6 subtype, also detected in 2007 (emm6.37), showed coresistance to 14- and 15-membered macrolides mediated by the mefA gene. Only three HR isolates were detected (isolates emm68.1/ST247/T3,13,B3264, emm77/ST399/T28, and emm28/ST52/T28), and all were identified in the retrospective study. Overall, the 673 isolates represented 25 emm types. All LR isolates were clustered into two emm types: emm6 (six emm6 subtypes) and emm75. All the 156 emm6 isolates had LR, harbored the Ser79/Ala mutation in the parC gene product, and had the same sequence type (ST), ST382. Most (21/33) of the emm75 isolates had LR, showed the Ser79/Phe plus Asp91/Asn double mutation in the parC gene product, and were ST150. The Asp91/Asn mutation by itself did not confer resistance to FQs.