PURPOSE: This study aimed to investigate the association between respiratory virus infection and pneumococcal colonization in children. METHODS: From May 2009 to June 2010, nasopharyngeal (NP) aspirates were obtained from patients under 18 years old who visited Seoul National University Children's Hospital for respiratory symptoms. NP samples were used to detect respiratory viruses (influenza virus A and B, parainfluenza virus 1, 2 and 3, respiratory syncytial virus A and B, adenovirus, rhinovirus A/B, human metapneumovirus, human coronavirus 229E/NL63 and OC43/HKU1) by RT-PCR and pneumococcus by culture. RESULTS: Median age of the patients was 27 months old. A total of 1,367 NP aspirates were tested for respiratory viruses and pneumococcus. Pneumococcus was isolated from 228 (16.7%) of samples and respiratory viruses were detected from 731 (53.5%). Common viruses were rhinovirus (18.4%), respiratory syncytial virus (RSV) A (10.6%), adenovirus (6.9%), influenza virus A (6.8%). Pneumococcal isolation rate was significantly higher in the cases of positive virus detection than negative detection [21.3% (156/731) vs. 11.3% (72/636), P<0.001]. For individual viruses, pneumococcal isolation rate was positively associated with detection of influenza virus A [24.7% (23/93) vs 16.1% (205/1274), P=0.001], RSV A [28.3% (41/145) vs 15.3% (187/1222), P=0.001], RSV B [31.3% (10/32) vs 16.3% (218/1335), P=0.042], rhinovirus A/B [22.6% (57/252) vs 15.3% (171/1115), P=0.010]. CONCLUSION: The study revealed that pneumococcal isolation from NP aspirates is related with respiratory virus detection. The result of this study could be used to investigate how respiratory viruses and pneumococcus cause clinical diseases.
Adenoviridae ; Child* ; Colon* ; Coronavirus ; Humans ; Metapneumovirus ; Orthomyxoviridae ; Paramyxoviridae Infections ; Respiratory Syncytial Viruses ; Rhinovirus ; Seoul ; Streptococcus pneumoniae

Adult ; Disease Outbreaks ; Humans ; Metapneumovirus ; Paramyxoviridae Infections ; epidemiology ; Respiratory Tract Infections ; epidemiology ; virology ; Students

Owing to advancements in molecular diagnostics, recent years have seen an increasing number of laboratories adopting respiratory viral panels to detect respiratory pathogens. In December 2015, the NxTAG respiratory pathogen panel (NxTAG RPP) was approved by the United States Food and Drug Administration. We compared the clinical performance of this new assay with that of the xTAG respiratory viral panel (xTAG RVP) FAST v2 using 142 clinical samples and 12 external quality assessment samples. Discordant results were resolved by using a laboratory-developed respiratory viral panel. The NxTAG RPP achieved 100% concordant negative results and 86.6% concordant positive results. It detected one coronavirus 229E and eight influenza A/H3N2 viruses that were missed by the xTAG RVP FAST v2. On the other hand, the NxTAG RPP missed one enterovirus/rhinovirus and one metapneumovirus that were detected by FAST v2. Both panels correctly identified all the pathogens in the 12 external quality assessment samples. Overall, the NxTAG RPP demonstrated good diagnostic performance. Of note, it was better able to subtype the influenza A/H3N2 viruses compared with the xTAG RVP FAST v2.
Coronavirus ; Hand ; Influenza, Human ; Metapneumovirus ; Pathology, Molecular ; Respiratory Tract Infections ; United States Food and Drug Administration

OBJECTIVETo determine whether human metapneumovirus (hMPV) was circulating in Suzhou area and the epidemiology and clinical features associated with hMPV infection.

METHODSamples were collected from January 2006 to December 2007; respiratory specimens were tested for the presence of hMPV by reverse-transcription polymerase Chain reaction (RT-PCR). PCR products of hMPV N gene from some patients were randomly selected for sequencing analysis, and the sequences of the nucleotides and deduced amino acids were compared with those in the GenBank.

RESULTOf the 4702 patients screened, 8% had evidence of hMPV infection. The positive rate in 2006 and 2007 was 8.4% and 7.6%, respectively. The positive rates detected during January to March, November and December were higher. The median age of patients was 22. 56 months. The infected children were diagnosed as having upper respiratory tract infection (3.2%), laryngitis (2.1%), bronchiolitis (27.1%), pneumonia (55.9%), and asthma exacerbation (11.7%). Sequence analysis of these hMPV N genes showed 99%-100% homology with the registered sequence in GenBank.

CONCLUSION(1) hMPV accounted for a significant proportion of respiratory tract infection in infants and children. (2) hMPV prevailed predominantly in the winter and spring time. (3) Clinically, hMPV infection can not be discriminated from the infection with other respiratory tract viruses.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Metapneumovirus ; Prevalence ; Respiratory Tract Infections ; epidemiology ; virology

PURPOSE:To identify the clinical features of human metapneumovirus (hMPV) and the respiratory syncytial virus (RSV) infection in children. METHODS:The participants of our study were 1,104 children who were admitted to Bungdang CHA hospital for lower respiratory infection from August 2006 through July 2007. Nasopharyngeal swabs were taken from the patients, and viruses were identified by RT-PCR. The clinical features of 51 patients with hMPV infection and 138 patients with RSV infection were compared by retrospective review of their medical records. RESULTS:The peak incidence of hMPV infection was noted in April, and that of RSV was noted in November. Both viruses had the highest incidence in patients age <1 year and hMPV infections occurred in 40% and 25% of patients age <6 months, respectively. In the hMPV group, 33.4% of the patients were age > or =2 years, while in the RSV group, 19% were age > or =2 years. In both groups, pneumonia was the most common clinical diagnosis, followed by acute bronchiolitis, acute bronchitis, and asthma. The white blood cell counts were higher in the RSV group, and fever was more frequent on admission in the hMPV group. CONCLUSION:hMPV was the fourth most common virus causing lower respiratory tract infections in children. The clinical features of hMPV infection were similar to those of RSV infection. This study may be helpful for the effective treatment of lower respiratory tract infection in children.
Asthma ; Bronchiolitis ; Bronchitis ; Child ; Fever ; Humans ; Incidence ; Leukocyte Count ; Metapneumovirus ; Pneumonia ; Respiratory Syncytial Viruses ; Respiratory Tract Infections ; Retrospective Studies ; Viruses

PURPOSE: This study was perfomed to analyze in detail the viral etiology of acute lower respiratory tract infections (ALRI) in Cheunan, Korea by multiplex RT-PCR, including human rhinovirus (hRV) and newly identified viruses such as human metapneumovirus (hMPV) and human coronavirus (HCoV-OC43, HCoV-229E/NL63). Method: Nasopharyngeal aspirates (NPA) were collected from 863 hospitalized children with ALRI on the first day of admission at Soonchunhyang University Cheonan Hospital and analyzed by multiplex RT-PCR from December 2005 to November 2006. RESULTS: Viral agents were detected from 474 subjects (54.9%). The identified viral pathogens were hRV 9.2%, hMPV 6.8%, HCoV-229E/NL63 1.4%, and HCoV-OC43 2.1%. Coinfections with > or =2 viruses were observed in 108 patients (22.8%). The major period of viral ALRI was the first year of life. Clinical diagnoses of viral ALRI were pneumonia (59.5%), bronchiolitis (24.7%), tracheobronchitis (11.4%), and croup (4%). The most common causes of bronchiolitis was respiratory syncytial virus B (RSV B), whereas hMPV, hRV, HCoV-229E/NL63, and HCoV-OC43 were commonly found in patients with pneumonia. The number of hMPV infections peaked between March and May 2006. HCoV-OC43 was prevalent from November to February 2006, whereas HCoV-229E and hRV were detected throughout the year. CONCLUSION: Although the study was confined to one year, hMPV was not detected during winter and peaked between March and April, which was not consistent with previous studies'. This present study indicates that HCoV is a less common respiratory pathogen in cases of ALRI in Korean children.
Bronchiolitis ; Child ; Child, Hospitalized ; Coinfection ; Coronavirus ; Croup ; Humans ; Korea ; Metapneumovirus ; Pneumonia ; Respiratory Syncytial Viruses ; Respiratory System ; Respiratory Tract Infections ; Rhinovirus

BACKGROUND: Multiplex PCR assay is a sensitive tool for the detection of various respiratory viruses. Seeplex RV12 ACE Detection (Seegene, Korea) assay is a multiplex RT-PCR assay for detection of 12 respiratory viruses. We had observed several cases with faint bands in the test. Those results were investigated in this study. METHODS: A total of 163 specimens were tested with Seeplex RV12 ACE Detection assay. The amplicons showing faint band in electrophoresis were reamplified and sequenced. RESULTS: A total of 99 viruses were detected in 80 specimens (49.1%). Twenty-four amplicons showed faint band in eletrophoresis. All of influenza virus A, parainfluenza viruses (PIV), coronavirus OC43, human metapneumovirus (HMPV) and adenovirus amplicons were reamplified, but 4 of 12 human rhinovirus amplicons were not reamplified. Sequences of reamplified PCR products showed homology of 95-99% to those of corresponding viruses in the National Center for Biotechnology Information database. CONCLUSIONS: Faint bands of influenza virus A, PIV-1, PIV-3, coronavirus OC43, HMPV and adenovirus in Seeplex RV12 ACE Detection assay are specific bands and seems to be weak positive results.
Adenoviridae ; Biotechnology ; Coronavirus ; Coronavirus OC43, Human ; Electrophoresis ; Humans ; Metapneumovirus ; Multiplex Polymerase Chain Reaction ; Orthomyxoviridae ; Paramyxoviridae Infections ; Polymerase Chain Reaction ; Rhinovirus

BACKGROUNDHuman metapneumovirus (hMPV) has recently been recognized as a notable respiratory pathogen in children. However, no isolation processes and only a limited understanding of hMPV epidemiology present in Chinese children are documented by far.

METHODSNasopharyngeal aspirates from 86 hospitalized children with acute respiratory tract infections from December 2004 to July 2005 were inoculated onto Vero-E6 cells for hMPV isolation. Total RNA was extracted from infected cells with a cytopathic effect and then subjected to a RT-PCR amplification of the N and F genes of the hMPV. Nucleotide sequences of amplified F gene products were examined using a variation analysis with the MegAlign program and the phylogenetic tree construction using the neighbor-joining algorithm with a Phylip package.

RESULTSSix strains of hMPV were isolated from the samples during winter, spring and summer. The most common symptoms were coughing and cyanosis, and the diagnoses were bronchiolitis, bronchopneumonia, infantile asthma, or upper respiratory tract infection. All isolates were in the A2 genetic sublineage and shared a high percentage of homology with the F gene in the nucleotide (99.8%-100%) and amino acid (99.3%-100%) sequences.

CONCLUSIONSThis report indicates that hMPV is an important viral agent for acute respiratory tract infections present in Chongqing, China. Knowledge of phylogeny and genes will benefit the studies on the treatment and prophylaxis of hMPV infection.

China ; Humans ; Infant ; Infant, Newborn ; Metapneumovirus ; classification ; genetics ; isolation & purification ; Paramyxoviridae Infections ; virology ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction

BACKGROUNDHuman metapneumovirus (hMPV) was discovered by scientists in the Netherlands as a novel respiratory virus in 2001 and had been found in children with acute respiratory tract infections (ARTI) in China. The objective of this study was to determine the importance of hMPV infection in children in Beijing and the genotypes of the circulating virus by the surveillance during a four-consecutive-year period.

METHODSClinical specimens collected from children with ARTI from January 2006 to December 2009 were tested for hMPV by RT-PCR using primers targeting the matrix (M) gene, followed by genotyping of hMPV directly from positive samples by diplex PCR with primers for glycoprotein (G) genes. Sequence analysis was used for genotyping of those un-typable samples. Common respiratory viruses in these clinical specimens were tested by virus isolation and antigen detection, in addition to hMPV detection.

RESULTSOf 4730 tested specimens, 191 (4.0%) were positive for hMPV and 62.8% of 191 were identified as genotype A. The positive rate of hMPV from hospitalized patients was higher than that from outpatients each year. Most of hMPV positive children were under five years old. The peak of hMPV activity mostly occurred in late spring and overlapped with or followed that of respiratory syncytial virus (RSV) and followed by parainfluenza virus 3. Of hMPV infected cases, 68.6% were lower respiratory tract infection, among which 79.4% were hospitalized, and upper respiratory tract infection was diagnosed for 31.4% of hMPV infected children. The 9.4% of hMPV positive samples were found to co-exist with other respiratory viruses.

CONCLUSIONShMPV was an important pathogen for ARTI in pediatric patients, especially those under five years old. Both genotypes A and B circulated simultaneously in Beijing.

Adolescent ; China ; Female ; Genotype ; Humans ; Male ; Metapneumovirus ; genetics ; pathogenicity ; Respiratory Tract Infections ; virology ; Reverse Transcriptase Polymerase Chain Reaction

Numerous studies have indicated that human metapneumovirus (hMPV) is an important viral pathogen in acute respiratory infections in children, presenting similar manifestations with respiratory syncytial virus (RSV). HMPV infection peaks in the winter-spring season and is more prevalent in younger ages, especially in children less than 1 year old. Host innate immune response has been implicated in recognition of pathogen-associated molecular patterns (PAMPs) of the virus. This recognition occurs through host pattern recognition receptors (PRRs). Toll like receptors (TLRs) are one of the largest class of PRRs which initiate and regulate adaptive immune responses. Some studies have indicated that TLR 3 and TLR 4 may play critical roles in hMPV infection. Construction of recombinant mutant viruses lacking one or two N-linked glycosylation sites in the F protein by using site-directed mutagenesis and reverse genetics may be helpful for developing attenuated live vaccines.
Humans ; Metapneumovirus ; immunology ; Paramyxoviridae Infections ; etiology ; prevention & control ; Vaccines, Attenuated ; immunology ; Vaccines, Synthetic ; immunology ; Viral Vaccines ; immunology