Molecular characterization of respiratory syncytial viruses infecting children reported to have received palivizumab immunoprophylaxis.

BACKGROUND: Respiratory syncytial virus (RSV) is a major cause of respiratory infections in children. Palivizumab (PZ) is the only RSV-specific immunoprophylaxis approved by the U.S. Food and Drug Administration. Mutations leading to amino acid substitutions in the PZ binding site of the RSV F protein have been associated with breakthrough RSV infections in patients receiving PZ. OBJECTIVE: To detect PZ resistance conferring mutations in RSV strains from children who received PZ. STUDY DESIGN: Children aged ≤ 24 months on October 31 who were hospitalized or had outpatient visits for respiratory illness and/or fever during October-May 2001-2008 in 3 US counties were included. PZ receipt was obtained from parent interviews and medical records among children subsequently infected with RSV. Archived nasal/throat swab specimens were tested for RSV by real-time RT-PCR. The coding region of the PZ binding site of the RSV F protein was sequenced using both Sanger and pyrosequencing methods. RESULTS: Of 8762 enrolled children, 375 (4.3%) were tested for RSV and had a history of PZ receipt, of which 56 (14.9%) were RSV-positive and 45 of these had available archived specimens. Molecular typing identified 42 partial F gene sequences in specimens from 39 children: 19 single RSV subgroup A, 17 subgroup B and 3 mixed infections. Nucleotide substitutions were identified in 12/42 (28.6%) RSV strains. PZ resistance mutations were identified in 4 (10.2%) of the 39 children, of which one had documented PZ receipt. CONCLUSIONS: Although RSV PZ resistance mutations were infrequent, most RSV-associated illnesses in children with a history of PZ receipt were not due to strain resistance.

High titers of IgE antibody to dust mite allergen and risk for wheezing among asthmatic children infected with rhinovirus.

BACKGROUND: The relevance of allergic sensitization, as judged by titers of serum IgE antibodies, to the risk of an asthma exacerbation caused by rhinovirus is unclear. OBJECTIVE: We sought to examine the prevalence of rhinovirus infections in relation to the atopic status of children treated for wheezing in Costa Rica, a country with an increased asthma burden. METHODS: The children enrolled (n= 287) were 7 through 12 years old. They included 96 with acute wheezing, 65 with stable asthma, and 126 nonasthmatic control subjects. PCR methods, including gene sequencing to identify rhinovirus strains, were used to identify viral pathogens in nasal washes. Results were examined in relation to wheezing, IgE, allergen-specific IgE antibody, and fraction of exhaled nitric oxide levels. RESULTS: Sixty-four percent of wheezing children compared with 13% of children with stable asthma and 13% of nonasthmatic control subjects had positive test results for rhinovirus (P< .001 for both comparisons). Among wheezing subjects, 75% of the rhinoviruses detected were group C strains. High titers of IgE antibodies to dust mite allergen (especially Dermatophagoides species) were common and correlated significantly with total IgE and fraction of exhaled nitric oxide levels. The greatest risk for wheezing was observed among children with titers of IgE antibodies to dust mite of 17.5 IU/mL or greater who tested positive for rhinovirus (odds ratio for wheezing, 31.5; 95% CI, 8.3-108; P< .001). CONCLUSIONS: High titers of IgE antibody to dust mite allergen were common and significantly increased the risk for acute wheezing provoked by rhinovirus among asthmatic children.

A naturally occurring human adenovirus type 7 variant with a 1743 bp deletion in the E3 cassette.

Human adenovirus type 7 (HAdV-7) is an important cause of acute respiratory disease (ARD). Different genomic variants of HAdV-7 have been described, designated 7a-7l. In a previous study to investigate risk factors for ARD and wheezing, nasopharyngeal samples were collected from 90 ill children seeking medical attention in Ribeirão Preto, São Paulo, Brazil. HAdVs were identified in 31 samples and were characterized by serum neutralization and genome restriction analysis. Eleven HAdVs were identified as being HAdV-7, five of which were classified as being of genome type 7p (Gomen). Six other HAdV-7 isolates gave new restriction profiles with all enzymes used and were classified as being a new genomic variant, 7m. These isolates were further characterized by sequencing. The hexon and fiber genes of the 7m variant were nearly identical to the prototype, 7p. However, nucleotide sequences from the E3 cassette revealed a 1743 bp deletion affecting the 16.1K, 19K, 20.1K and 20.5K ORFs.

Human bocavirus species 2 and 3 in Brazil.

BACKGROUND: The newly described human bocavirus (HBoV) species 2 and 3 have been repeatedly detected in stool strengthening the possibility that these viruses might present a tropism for the gastrointestinal tract and may be etiological agents of diarrhea. OBJECTIVE: In this study we assessed the presence of HBoV2 and HBoV3 in stool specimens from Brazilians with acute gastroenteritis. STUDY DESIGN: Stool samples from Brazilian patients with acute diarrhea were analyzed for HBoV2 and HBoV3 by PCR assay. Full or partial genome sequences were obtained for selected isolates. Electron microscopy analysis was used to investigate virus morphology. RESULTS: Electron microscopy confirmed the presence of virus-like particles in HBoV PCR-positive specimens, with morphology similar to other members of the Parvoviridae family. Five samples out of 807 (0.6%) were positive for HBoV3. Three of the HBoV3-positive patients were HIV/AIDS positive. A selected group of 144 samples was also tested for HBoV2 and 30 samples (20.8%) were positive, 11 of which were HIV/AIDS positive. CONCLUSION: This study reports the detection and genetic characterization of HBoV3 and HBoV2 in the stool of Brazilian patients with acute diarrhea. This is the first description of HBoV3 outside Australia, suggesting a wide global distribution of this virus. Further studies are needed to better understand the role of HBoV in gastrointestinal infections, particularly among patients with HIV/AIDS.

Parainfluenza virus infection of young children: estimates of the population-based burden of hospitalization.

OBJECTIVE: To determine the population-based inpatient disease burden of parainfluenza virus in children <5 years of age. STUDY DESIGN: The New Vaccine Surveillance Network (NVSN) enrolled children <5 years of age who were hospitalized with febrile or acute respiratory illnesses. Surveillance hospitals admitted >95% of all hospitalized children from each county. Combined nasal turbinate/throat swabs were tested for parainfluenza virus (PIV), respiratory syncytial virus, and influenza virus with culture and reverse-transcription-polymerase chain reaction. Both parental interviews and medical chart reviews were conducted. Age-specific population-based hospitalization rates were calculated. RESULTS: From October 2000 through September 2004, 2798 children were enrolled. A total of 191 PIVs were identified from 189 children (6.8% of enrolled: 73 PIV type 1, 23 PIV type 2, and 95 PIV type 3), compared with 521 respiratory syncytial viruses and 159 influenza viruses. Mean PIV hospitalization rates were 3.01, 1.73, 1.53, 0.39, and 1.02 per 1000 children per year for ages 0 to 5 months, 6 to 11 months, 12 to 23 months, 24 to 59 months, and 0 to 59 months, respectively. CONCLUSIONS: PIV accounted for 6.8% of all hospitalizations for fever, acute respiratory illnesses, or both in children <5 years of age. The pediatric PIV inpatient burden is substantial and highlights the need to find an effective vaccine candidate.

Surveillance of eight respiratory viruses in clinical samples of pediatric patients in southeast Brazil.

OBJECTIVE: Detection of the eight most common respiratory viruses: human respiratory syncytial virus (HRSV), influenza virus A and B (IA and IB), parainfluenza viruses 1, 2 and 3 (HPIV1, 2 and 3), adenovirus (Ad) and human metapneumovirus (HMPV), in order to establish the etiology of acute respiratory infections (ARIs) and the epidemiology of these viruses in young children seen at Hospital Universitário, Universidade de São Paulo, in São Paulo, Brazil, during 2003. METHODS: The epidemiological surveillance was conducted in all children younger than 5 years hospitalized at the Hospital for lower respiratory tract infections (LRTI) from January 1, 2003 to December 30, 2003. Nasal and throat samples were scanned for respiratory viruses by polymerase chain reaction and detected by the GeneScan assay. RESULTS: Of 336 samples collected from 336 patients, 187 (55.6%) were positive for at least one of the respiratory viruses studied. Of all the children, HRSV was identified in 24.1%, HMPV in 17.8%, HPIV3 in 8.3%, Ad in 6.8%, IA in 5%, HPIV1 in 0.6%, but no virus could be detected in 44.1%. Dual virus infections were detected in 7.1% of all samples (12.8% of positive samples). HPIV2 and IB were not detected in the present study. CONCLUSIONS: This study confirms that children younger than 5 years and particularly younger than 1 year have a high hospitalization rate due to HRSV, HMPV, HPIV, influenza and adenovirus. We were able to determine the etiology and epidemiology of most ARIs and trace the seasonal profile of the commonest respiratory viruses among young children.

Vigilância de oito vírus respiratórios em amostras clínicas de pacientes pediátricos no sudeste do Brasil / Surveillance of eight respiratory viruses in clinical samples of pediatric patients in Southeast Brazil

OBJETIVO: Detecção de oito vírus respiratórios mais comuns: vírus respiratório sincicial humano (VRSH), vírus influenza tipo A e B (IA e IB), vírus da parainfluenza 1, 2 e 3 (VPIH1, 2 e 3), adenovírus (Ad) e metapneumovírus humano (MPVH), a fim de estabelecer a etiologia das infecções respiratórias agudas (IRA) e a epidemiologia desses vírus em crianças pequenas atendidas no Hospital Universitário da Universidade de São Paulo, em São Paulo, Brasil, durante o ano de 2003. MÉTODOS: A vigilância epidemiológica foi realizada em todas as crianças menores de 5 anos hospitalizadas por causa de doenças do trato respiratório inferior (DTRI) entre 1º de janeiro de 2003 e 20 de dezembro de 2003, no hospital universitário. Amostras coletadas de nasofaringe foram analisadas quanto à presença de vírus respiratórios através da reação em cadeia da polimerase e detectadas pelo programa GeneScan. RESULTADOS: Das 336 amostras coletadas, 187 (55,6 por cento) foram positivas para pelo menos um dos vírus respiratórios estudados. De todas as crianças, o VRSH foi identificado em 24,1 por cento, o MPVH em 17,8 por cento, o VPIH3 em 8,3 por cento, o Ad em 6,8 por cento, o IA em 5 por cento, o VPIH1 em 0,6 por cento, sendo que nenhum vírus foi detectado em 44,1 por cento. Infecções virais duplas foram detectadas em 7,1 por cento de todas as amostras (12,8 por cento das amostras positivas). O VPIH2 e o IB não foram detectados no presente estudo. CONCLUSÕES: Este estudo confirma que as crianças menores de 5 anos, e especialmente aquelas menores de 1 ano, apresentam uma alta taxa de hospitalização devido aos seguintes vírus: VRSH, MPVH, VPIH, influenza e adenovírus. Foi possível determinar a etiologia e epidemiologia da maioria das IRAs e traçar o perfil de sazonalidade dos vírus respiratórios mais comuns entre as crianças pequenas.

OBJECTIVE: Detection of the eight most common respiratory viruses: Human respiratory syncytial virus (HRSV), influenza virus A and B (IA and IB), parainfluenza viruses 1, 2 and 3 (HPIV1, 2 and 3), adenovirus (Ad) and human metapneumovirus (HMPV), in order to establish the etiology of acute respiratory infections (ARIs) and the epidemiology of these viruses in young children seen at Hospital Universitário, Universidade de São Paulo, in São Paulo, Brazil, during 2003. METHODS: The epidemiological surveillance was conducted in all children younger than 5 years hospitalized at the Hospital for lower respiratory tract infections (LRTI) from January 1, 2003 to December 30, 2003. Nasal and throat samples were scanned for respiratory viruses by polymerase chain reaction and detected by the GeneScan assay. RESULTS: Of 336 samples collected from 336 patients, 187 (55.6 percent) were positive for at least one of the respiratory viruses studied. Of all the children, HRSV was identified in 24.1 percent, HMPV in 17.8 percent, HPIV3 in 8.3 percent, Ad in 6.8 percent, IA in 5 percent, HPIV1 in 0.6 percent, but no virus could be detected in 44.1 percent. Dual virus infections were detected in 7.1 percent of all samples (12.8 percent of positive samples). HPIV2 and IB were not detected in the present study. CONCLUSIONS: This study confirms that children younger than 5 years and particularly younger than 1 year have a high hospitalization rate due to HRSV, HMPV, HPIV, influenza and adenovirus. We were able to determine the etiology and epidemiology of most ARIs and trace the seasonal profile of the commonest respiratory viruses among young children.