南京地区急性呼吸道感染患儿人博卡病毒的检出与分析

目的 了解南京地区急性呼吸道感染患儿HBoV的检出情况,并探讨HBoV与临床特征的关系.方法 收集2009年7月至2010年6月南京医科大学附属南京儿童医院呼吸科收治的397例急性呼吸道感染住院患儿为病例组,对照组为50例无呼吸道感染症状的儿童.应用实时荧光定量PCR法检测鼻咽分泌物标本HBoV.对HBoV阳性标本,应用实时荧光定量PCR法检测MP和CT,直接免疫荧光法检测RSV、ADV、IVA、IVB、PIV-1、PIV-2、PIV-3和hMPV.随机选取5份HBoV阳性标本扩增HBoV NP-1片段后进行核苷酸序列测定,结果与GenBank中已知序列进行比对,绘制系统进化树.结合HBoV阳性患儿临床资料,分析HBoV的流行病学特点、临床表现和最终临床诊断.结果 实时荧光定量PCR法检出HBoV DNA阳性率为8.3%(33/397),其中有57.6%(19/33)混合其他病原体感染.与HBoV混合感染的前3位病原体依次为MP(27.3%,9/33)、RSV(24.2%,8/33)和PIV.3(12.1%,4/33).7～36个月龄感染HBoV患儿有25例,占HBoV DNA阳性患儿的75.8%(25/33).5份标本HBoV NP-1基因序列均一致,与st1、st2和WHL-1等序列同源性为99%～100%.结论 HBoV是南京地区急性呼吸道感染患儿的病原之一.HBoV NP-1基因高度保守,在不同地区和不同时期的流行株间变异较小,可作为实时荧光定量PCR等方法的检测标记.

Abstract：

Objective To investigate the possible existence of HBoV in children with acute respiratory infections in Nanjing area and explore its relationship with clinical characteristics.Methods A total of 397 nasopharyngeal secretion samples were collected from children with acute respiratory infection,admitted from July 2009 to June 2010 in Nanjing Children'S Hospital affiliated to Nanjing Medical University,and 50 cases of children without symptoms of respiratory infection were recruited as control group,whose nasopharyngeal secretion samples were also collected.HBoV was determined by real-time fluorescence quantitative PCR.MP and CT were detected by real-time fluorescence quantitative PCR in those HBoV-positive samples.RSV,ADV,IVA,IVB,PIV-1,PIV-2,PIV-3 and hMPV were detected by direct antigen-specific immunofluorescence assays.HBoV NP-1 fragments were amplified and sequenced in 5 HBoV positive samples randomly selected.The results were compared with the known GenBank sequence,and thereby the phylogenetic tree was established.The epidemiological characteristics,clinical presentation and the final clinical diagnosis of HBoV were analyzed according to the clinical data of the HBoV-positive patients.Results Thirty-three HBoV-positive cases were detected by real-time fluorescence quantitative PCR method with a positivity rate of 8. 3% ( 33/397 ). Among the 33 HBoV-positive cases, 19 cases (57.6%) were multiple infections with HBoV and other pathogens, the top three of which were MP (27.3% ,9/33 ),RSV (24.2% , 8/33 ) and PIV-3 ( 12. 1% ,4/33 ). Affected children aged from 7 to 36 months old accounted for 75.8% of the total ( 25/33 ). The measured HBoV NP-1 gene sequences of 5 specimens were consistent,indicating a high homology (99% to 100% ) with the stl, st2 and WHL-1. Conclusions HBoV is one of the pathogens of children's acute respiratory infections in Nanjing. HBoV NP-1 gene is highly conserved,with little variation in different seasons and in different regions and therefore can be used as a marker for real-time fluorescence quantitative PCR and other methods.

Detection and analysis of human bocavirus in children with acute respiratory infection in Nanjing

Objective To investigate the possible existence of HBoV in children with acute respiratory infections in Nanjing area and explore its relationship with clinical characteristics.Methods A total of 397 nasopharyngeal secretion samples were collected from children with acute respiratory infection,admitted from July 2009 to June 2010 in Nanjing Children'S Hospital affiliated to Nanjing Medical University,and 50 cases of children without symptoms of respiratory infection were recruited as control group,whose nasopharyngeal secretion samples were also collected.HBoV was determined by real-time fluorescence quantitative PCR.MP and CT were detected by real-time fluorescence quantitative PCR in those HBoV-positive samples.RSV,ADV,IVA,IVB,PIV-1,PIV-2,PIV-3 and hMPV were detected by direct antigen-specific immunofluorescence assays.HBoV NP-1 fragments were amplified and sequenced in 5 HBoV positive samples randomly selected.The results were compared with the known GenBank sequence,and thereby the phylogenetic tree was established.The epidemiological characteristics,clinical presentation and the final clinical diagnosis of HBoV were analyzed according to the clinical data of the HBoV-positive patients.Results Thirty-three HBoV-positive cases were detected by real-time fluorescence quantitative PCR method with a positivity rate of 8. 3% ( 33/397 ). Among the 33 HBoV-positive cases, 19 cases (57.6%) were multiple infections with HBoV and other pathogens, the top three of which were MP (27.3% ,9/33 ),RSV (24.2% , 8/33 ) and PIV-3 ( 12. 1% ,4/33 ). Affected children aged from 7 to 36 months old accounted for 75.8% of the total ( 25/33 ). The measured HBoV NP-1 gene sequences of 5 specimens were consistent,indicating a high homology (99% to 100% ) with the stl, st2 and WHL-1. Conclusions HBoV is one of the pathogens of children's acute respiratory infections in Nanjing. HBoV NP-1 gene is highly conserved,with little variation in different seasons and in different regions and therefore can be used as a marker for real-time fluorescence quantitative PCR and other methods.