【摘要】 目的　了解北京地区婴幼儿急性呼吸道感染与人类偏肺病毒 (HumanMetapneumovirus,HMPV)的关系。 方法　对 2 0 0 2年 1 1月～ 2 0 0 3年 3月 ,收集的 2 4 7份经过间接免疫荧光法和病毒分离排除了常见的呼吸道病毒感染的鼻咽洗液标本进行了HMPV基因的检测。用针对HMPVN基因序列设计的特异引物对这 2 4 7份标本进行RT PCR扩增 ,扩增片段经 1 2 %琼脂糖凝胶电泳检测。挑选 1 0份阳性扩增产物直接进行核苷酸序列测定 ,并将所测序列在GeneBank中进行比较分析。结果　 2 4 7份标本中共检测到 74份RT PCR阳性扩增产物 ,阳性率为 30 0 % (74/2 4 7)。 74例中临床诊断为肺炎者 36例 ,占 48 6 % ;其次为毛细支气管炎 2 1例 ,占 2 8 4% ;年龄 2个月～ 5岁 7个月不等 ,≤ 2岁者占 83 8%。所挑选的 1 0份N基因扩增产物与GeneBank中同源性最高的均为HMPV(87%～ 99% )。 1 0份标本中 9份标本的扩增产物之间的核苷酸和氨基酸同源性在95 8%～ 1 0 0 %和 98 6 %～ 1 0 0 % ,1份标本的扩增产物与其他标本同源性较低 ,仅为 87 3 %～ 89 7%和 95 8%～ 97 2 %。结论　 (1 )北京地区部分儿科患者的急性呼吸道感染与HMPV有关 ;(2 )北京地区的HMPV中可能存在不同的基因型更多还原

【Abstract】 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 immunofluoresence 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 GeneBank 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 GeneBank, 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% Conclusion 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更多还原