2009甲型H1N1流感流行期间北京急性呼吸道感染儿童常见呼吸道病毒的研究

摘要

目的 了解2009至2010年甲型H1N1流感大流行期间北京地区急性呼吸道感染患儿中,除流感病毒外的其他几种常见呼吸道病毒感染的流行情况.方法 设计并建立检测包括呼吸道合胞病毒(RSV)A/B亚型,副流感病毒(PIV)1、2、3型,腺病毒(ADV)和人博卡病毒(HBoV)在内的多重实时荧光PCR方法,并应用该方法 对2009年6月至2010年2月甲型H1N1流感大流行期间,对首都儿科研究所附属儿童医院就诊的急性呼吸道感染患儿中甲型H1N1流感病毒检测阴性的咽拭子标本进行上述呼吸道病毒的核酸检测.结果 新建立的多重RT-PCR方法 最低可检测的目标基因含量为10~300拷贝数,未见与其他非目标病毒的交叉反应.本研究共检测标本849份,病毒检测总阳性率为39.0%(331份),5岁以下占87.6%(290/331份).各病毒的检测阳性率分别为RSV-A亚型1.4%(12份),RSV-B亚型8.4%(71份),PIV-1型8.2%(70份),PIV-2型0.5%(4份),PIV-3型3.9%(33份),ADV 13.9%(118份),HBoV 2.7%(23份).RSV检出以B亚型为主(85.5%),流行高峰在2009年11月至2010年2月.PIV检出以PIV-1型及PIV-3型为主,PIV-1型的流行高峰在2009年7月至2009年10月,PIV-2型及PIV-3型的流行高峰在2009年6～9月.ADV病毒在研究期间均有较高检出率(平均为13.9%),HBoV的流行高峰在2009年9～12月.结论 除流感病毒外,ADV、RSV-B及PIV可能也是2009甲型H1N1流感流行期间引起儿童急性呼吸道感染的重要病原.比较以往的资料可见各病原在2009甲型H1N1流感流行期间的流行季节性及检出率基本未受到新型流感病毒的影响.%Objective To investigate the prevalence of common respiratory viruses in infants and young children with acute respiratory infection (ARI) during the pandemic of 2009 influenza A( H1N1 ) in Beijing. Methods Primers and probes were designed to develop a novel multiplex real-time PCR for the detection of 7 respiratory viruses including respiratory syncytial virus ( RSV ) types A and B, parainfluenza virus ( PIV ) types 1, 2 and 3, adenovirus ( ADV ) and human bocavirus( HBoV ) in clinical samples. Clinical specimens were collected from children who visited the Children's Hospital affiliated to Capital Institute of Pediatrics with ARI from June 2009 to February 2010 and had been tested negative for 2009 influenza A( H1N1 ). Results The method could detect as few as 10-300 copies of each target plasmid DNA, and could only specifically detect the target virus but not other viruses in specimens. The application of the method in 849 throat swab specimens indicated that the overall positive rate was 39.0%, 1.4% for RSV-A, 8.4% for RSV-B, 8.2% for PIV-1, 0.5% for PIV-2, 3.9% for PIV-3, 13.9% for ADV, 2.7% for HBoV, respectively. In the tested positive cases, 87. 6% cases were under 5 years of age. Statistical analysis of the data demonstrated that the dominated subtype of RSV was RSV-B ( 85. 5% ) which caused an epidemic peak from Noy. 2009 to Feb. 2010; PIV infections were mainly associated with PIV-1 from Jul. 2009 to Oct. 2009 and with PIV-3 from Jun. 2009 to Sep. 2009 ; ADV had a relatively high detectable rate during the whole period ( an average of 13. 9% ); and the detectable rate of HBoV increased from Sep. 2009 to Dec. 2009. Conclusions In addition to influenza virus, ADV, RSV-B and PIV infections were also associated with ARI in children during 2009 pandemic influenza A( H1N1 )in Beijing. No evidence showed that the characteristics of both the seasonality and the morbidity of the viruses involving in this study were impacted by the pandemic of the new influenza virus H1N1.