[Human metapneumovirus may associate with acute respiratory infections in hospitalized pediatric patients in Beijing, China].
OBJECTIVE
A new respiratory virus, human metapneumovirus (HMPV) was recently identified by scientists in the Netherlands first and then in a few other countries. To investigate if this newly discovered virus is associated with the acute respiratory infections in pediatric patients in Beijing, tests were developed to detect HPMV gene fragments from nasopharyngeal aspirates collected from infants and young children hospitalized for acute respiratory infections from November 2002 to March 2003.


METHODS
The HMPV was screened by reverse transcription-polymerase chain reaction (RT-PCR). RNAs were extracted by Trizol from 247 specimens which had been determined as negative for conventional respiratory viruses including RSV, influenza A and B, parainfluenza I, II, III and adenovirus by indirect immunofluorescence test as well as virus isolation. The HMPV RNAs were detected by reverse transcription tests using random primer and M-MLV reverse transcriptase followed by PCR using the primers designed from the published sequence of the N protein-encoding gene from the first HMPV identified in the Netherlands. PCR products were visualized by 1.2% agarose gel electrophoresis. Selected positive PCR products were sequenced and the sequences of the nucleotides and deduced amino acids were compared with those in the GenBank.


RESULTS
Among those 247 specimens negative for common respiratory viruses, 74 (30.0%) showed the predicted 213 bp PCR products in agarose gel. Most of clinical diagnoses for these 58 patients were pneumonia (36, 48.6%), bronchiolitis (21, 28.4%), and bronchitis and asthma in some patients. Nearly 90 percent of positive specimens were from patients under 2 years of age. Ten out of 74 amplicons were randomly selected for sequence analysis. When compared with the sequences in the GenBank, the nucleotide sequences of these 10 amplicons shared high homology only with those of HMPVs. The nucleotide sequence identities of these 10 samples with those from the Netherlands and Canada were 87% - 99%. When compared with the nucleotide sequence from the first reported strain by Van den Hoogen (strain HMPV 00-1), the sequence identities of these 10 fragments ranged from 88.7% to 99.1%. Among the 10 amplicons from the specimens, the nucleotide identities were 87.3% - 100%. One of the 10 amplicons (No. 1816) shared lower identity with others (87.3% - 89.7%), whereas the other 9 shared higher identities (95.8% - 100%) with each other. The comparison of amino acids showed that these 10 amplicons showed high homology (95.8% - 100%). Again, amplicon No.1816 shared lower homology (95.8% - 97.2%) with others, whereas the other 9 shared higher homology (98.6% - 100%). The amino acid homology between No.1816 and HMPV 00-1 was 95.8%, whereas that of the other 9 with HMPV 00-1 was 98.6% - 100%.


CONCLUSIONS
These data suggested that some of acute respiratory infections in pediatric patients in Beijing area are related to the newly identified human metapneumovirus. The HMPV circulating in Beijing may have different genotypes.
Zhu
Ru-nan
RN
Laboratory of Virology, Beijing Municipal Laboratory of Infection and Immunity, and Children's Hospital Affiliated to Capital Institute of Pediatrics, Beijing,100020, China.
Qian
Yuan
Y
Deng
Jie
J
Wang
Fang
F
Hu
Ai-zhong
AZ
Lu
Jing
J
Cao
Li
L
Yuan
Yi
Y
Cheng
Hui-zhong
HZ
D000208:Acute Disease; D002648:Child; D002675:Child, Preschool; D002681:China; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007223:Infant; D008297:Male; D029121:Metapneumovirus / Q000235:genetics*; D019590:Nucleocapsid Proteins / Q000235:genetics; D018184:Paramyxoviridae Infections / Q000473:pathology / Q000821:virology*; D012367:RNA, Viral / Q000235:genetics; D012141:Respiratory Tract Infections / Q000473:pathology / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D019590:Nucleocapsid Proteins; D012367:RNA, Viral