2016年武汉地区住院儿童人博卡病毒感染的临床特征及流行病学

目的了解武汉地区住院儿童呼吸道人博卡病毒（HBoV）感染情况、临床特征及流行病学特点。方法收集2016年1—12月该院儿科因急性呼吸道感染的住院患儿968例,采用无菌负压吸引法采集新鲜标本检查痰HBoV、RSV、ADV、INF A/B和PIV I/II/III,分析HBoV感染病例的流行情况及临床特征。结果968例患儿中,HBoV阳性75例,检出率7.75%。男女患儿检出率分别为5.68%、2.07%,不同性别HBoV检出率比较,差异无统计学意义（χ2=2.083,P=0.140）;53例（70.67%）HBoV阳性患儿年龄＜1岁,63例（84.00%）HBoV阳性患儿年龄＜3岁,各年龄组检出率比较,差异有统计学意义（χ2=4.60,P=0.043）。春、夏、秋、冬HBoV检出率分别为9.78%、5.29%、2.86%、5.20%;除9、10月份外,HBoV在其余月份均有检出,3月份检出率最高（为33.33%）。17例单纯HBoV感染,58例混合感染,HBoV临床诊断以支气管肺炎为主（26例,占34.67%）。HBoV病例主要临床特征为发热（81.33%）、咳嗽和喘息（各占77.33％）。结论HBoV是武汉地区住院儿童呼吸道感染的重要病原体之一,全年均可发生HBoV感染,流行高峰在春季和夏季,无性别差异。住院儿童感染HBoV临床诊断以支气管肺炎最常见,主要表现为发热、咳嗽、喘息。     

人类博卡病毒的检出及病原学初步研究

目的 了解粤东地区喘息性疾病患儿的感染病原中是否存在人类博卡病毒(HBoV),并探讨其感染的临床特征.方法 收集急性喘息性疾病患儿鼻咽抽吸物(NPA)246份和健康体检儿童的NPA 83份,应用针对HBoV的核衣壳蛋白基因(VP1/VP2基因)的PCR引物进行HBOV基因片段检测.随机选取HBoV基因检测阳性样本中的7份,对其PCR扩增产物直接进行核苷酸序列测定,将所测到的序列与GenBank中的基因序列进行比较分析.同时,对HBoV PCR阳性病例临床资料进行分析.结果 246份样本中HBoV基因检测阳性的有13份,阳性率为5.8%(13/246);HBoV检测阳性样本主要分布于＜1岁的患儿,占阳性病例的72.9%(10/13),尤其是＜5个月者占阳性病例的53.8%(7/13).基因序列分析表明,本研究中7份HBoV之间基因序列的同源性在99.7%～100%之间;与WLL-1株相应片段的核苷酸同源性为98.1%～100%,与HBoV st1株的同源性为97.6%～98.5%,与HBoV st2株的同源性为98.1%～99.8%.相应的患儿均表现咳嗽、呼吸困难及喘息等临床症状,除1例诊断为哮喘急性发作外,其余为喘息性肺炎(5例)和毛细支气管炎(7例).83份健康体检儿童的呼吸道样本中均未检测到HBoV特异性基因片段.结论 粤东地区儿童急性呼吸道感染引起的喘息性疾病中,有部分系HBoV感染引起;且HBoV感染在低年龄组儿童常见,其临床表现与普通的喘息性肺炎和毛细支气管炎类似.     

天津地区儿童人博卡病毒感染及基因型分析

目的 了解天津地区急性下呼吸道感染儿童中人博卡病毒(HBoV)的感染情况及病毒基因特性.方法 采集2008年9月- 2009年5月急性下呼吸道感染住院儿童的鼻咽吸取物标本202份,进行PCR扩增来检测HBoVNP-1基因,对VP1及VP2基因进行扩增及序列测定,与GenBank中登录的序列进行比较,分析序列特征并绘制种系发生树,同时采用多重PCR方法检测其他12种呼吸道病毒及WU多瘤病毒.结果 202份标本中,6.9％( 14/202)为HBoV阳性,所有阳性患儿的年龄均≤6岁,6月～12月龄患儿感染率最高,为14.3％ (6/42);所有感染者均为肺炎和喘息性支气管炎患儿,各月份均有检出;HBoV与其他病毒的混合感染率为85.7％ (12/14),其中58.3％ (7/12)为3种以上呼吸道病毒混合感染;VP1/VP2基因种系发生树分析表明13株HBoV均属于Ib簇,同源性分析表明,天津地区HBoV VP1基因与从GenBank下载的Ⅰ型序列比较,核酸及氨基酸的同源性分别为99.0％～ 99.8％及99.0％ ～99.9％.结论 天津地区存在HBoV流行,感染者多为＜6岁儿童,HBoV与其他呼吸道病毒混合感染常见,HBoV病毒均属于Ib簇.     

杭州地区急性呼吸道感染患儿人博卡病毒的检测及分析

目的了解杭州地区人博卡病毒(HBoV)的流行情况和基因变异情况。方法对杭州地区160例儿童急性呼吸道感染患者咽拭子样本进行HBoV检测,并对阳性样本进行常见呼吸道病毒的检测,最终选取HBoV单一阳性的HZ1株和HZ2株进行VP1基因扩增,将获得的序列上传Genbank并与其他国家和地区的8株HBoV的VP1基因进行序列比对和分析。结果杭州地区HBoV的检出率为6.9%(11/160),与其他常见呼吸道病毒的合并感染率达到45.5%(5/11),HZ1株和HZ2株与其他国家和地区的8株HBoV的VP1基因相似性达到99%以上。结论研究证明杭州地区急性呼吸道感染患儿中存在HBoV的感染,与其他呼吸道病毒有较高的合并感染,从分离到的HZ1株和HZ2株来看,杭州地区流行的是ST2基因型,其外壳蛋白VP1基因变异很小。     

苏州地区人博卡病毒在儿童急性呼吸道感染中的流行特征

目的分析人博卡病毒(human bocavirus,HBoV)在苏州地区儿童急性呼吸道感染患儿中的流行特征。方法收集6108例急性呼吸道感染的住院儿童鼻咽分泌物标本,实时荧光PCR法检测HBoV,同时采用实时荧光定量PCR检测肺炎支原体(MP),直接免疫荧光法检测呼吸道合胞病毒(RSV)、流感病毒A型、B型(IVA,IVB)、副流感病毒1~3型(PIV1~3)和腺病毒(ADV)。结果共检出HBoV感染378例(6.19%,378/6108),其中HBoV与其他病原体混合感染73例(19.31%,73/378)。与HBoV混合感染的前三位病原体分别为RSV(64.38%,47/73),PIV-3(16.44%,12/73),MP(12.33%,9/73)。HBoV感染阳性率在夏秋季分别为9.06%和11.35%,而在冬春季3.52%和2.37%,夏秋和冬春各季之间阳性率存在显著性差异(均P<0.01)。HBoV感染在7~12月和1~3岁儿童中的阳性率分别为11.14%和10.40%,而在0~6月组、3~5岁组、5~6岁组和≥6岁组的阳性率分别为2.83%、4.29%、3.11%和1.97%,7月~3岁患儿与其它年龄组间HBoV阳性率存在显著性差异(χ2=144.26,P<0.01)。结论人博卡病毒是苏州地区儿童急性呼吸道感染的主要病原之一,苏州全年均存在HBoV感染,以夏秋季高发。HBoV易与其它病原混合感染,尤其是RSV。HBoV感染在7月~3岁儿童中高发。     

小儿急性呼吸道感染与人博卡病毒相关性的研究

目的:了解小儿急性呼吸道感染与人博卡病毒(Human Bocavirus,HBoV)的相关性。方法:对2008年1～5月在长春市儿童医院住院的1007例急性呼吸道感染患儿鼻咽深部分泌物样本用PCR扩增的方法进行HboV NS1基因检测。随机选取8例HBoV阳性产物进行测序,并将测到的序列与GenBank中的基因序列和HBoV阳性病例的临床资料进行分析。结果:1007例标本共检测出44例HBoV阳性,阳性率为4.37%,其中急性上呼吸道感染、急性支气管炎、毛细支气管炎和支气管肺炎患儿人博卡病毒阳性检出率分别为1.29%(2/155)、5.71%(2/35)、8.70%(2/23)和4.79%(38/794)。HboV急性呼吸道感染的年龄分布为1岁以下39例,占88.64%;1岁以上5例,占11.36%;男女性别比值是1.75∶1;临床表现咳嗽伴喘憋占97.56%,咳嗽伴腹泻占34.14%,入院时体温升高占54.55%,肺部听诊闻及干和/或湿性罗音者占95.12%,X线胸片片状影占87.80%,病程平均为13.79天。结论:人博卡病毒是引起小儿急性呼吸道感染的病原之一;HboV所致呼吸道感染部位多在下呼吸道;感染患儿以1岁以内婴儿为主。     

武汉地区人类博卡病毒呼吸道感染患儿的临床特点

目的研究武汉地区住院患儿呼吸道感染人类博卡病毒(Human Bocavirus,HBoV)的临床特点。方法选择2017年1-12月武汉市第三医院因呼吸道感染住院患儿1 438例以及健康儿童310名为研究对象。应用RT-PCR检测1 438例患儿咽拭子病毒感染情况;比较健康儿童和HBoV阳性患儿炎症指标水平;分析HBoV阳性患儿临床症状;采集HBoV阳性患儿咽拭子,进行扩增后基因测序,分析基因突变情况。结果 1 438例呼吸道感染住院患儿中HBoV阳性333例(23.16%)其中1～5岁患儿HBoV阳性率为46.4%;与对照组健康儿童相比,HBoV阳性患儿白细胞(WBC)、中性粒细胞计数(NEUT)、C-反应蛋白(CRP)、肿瘤坏死因子-α(TNF-α)水平显著升高,差异具有统计学意义(P0.05); 333例HBoV阳性患儿,其中高载量病毒(10~4 copies/ml)224例,低载量病毒109例,高载量与低载量病毒患儿的呼吸道症状无明显差别,但高载量病毒患儿更容易出现腹泻(P=0.016),更容易引起肺炎(P=0.034); 333例HBOV阳性临床标本基因测序,共获得48个全基因序列,其中15个为NP1基因序列,NP1基因236位点(G236A)、447位点(A447G)突变的样本中,病毒载量显著低于未突变的样本,差异具有统计学意义(P0.05)。结论呼吸道感染患儿中,HBoV阳性率较高,感染患儿易引起肺炎,炎症指标水平升高。某些位点基因突变可能影响病毒基因复制。     

免疫荧光法研究桐乡地区婴幼儿呼吸道偏肺病毒感染

目的研究桐乡地区婴幼儿急性呼吸道感染患者中偏肺病毒(hMPV)感染及临床意义。方法采用免疫荧光法,对2008年8月-2009年7月收集的431例急性呼吸道感染婴幼儿的呼吸道分泌物标本进行hMPV抗原检测;同时检测呼吸道合胞病毒(RSV)等其他常见呼吸道病毒的感染情况。结果 431份标本中检测到hMPV阳性68例,阳性率为15.78%;68例中临床诊断为肺炎者38例,占55.88%。结论 hMPV为桐乡地区婴幼儿急性呼吸道感染的重要病原体,冬季高发,春秋两季发病率也较高;hMPV感染易发展为肺炎等较重病症,在临床上应引起高度重视。     

2014-2015年武汉地区4 232例儿童呼吸道人类博卡病毒感染流行病学特征

目的 分析武汉地区4 232例急性呼吸道感染(ARTI)的儿童人类博卡病毒(HBoV)感染的临床及流行病学特征。 方法 收集2014年1月1日-2015年12月31日期间4 232例7岁以下儿童经确诊为ARTI的鼻咽抽吸物(NPA),利用RT-PCR法对HBoV进行筛查,同时对流感病毒A型(Inf-A)、流感病毒B型(Inf-B)、呼吸道合胞病毒(RSV)、腺病毒(ADV)及副流感病毒Ⅰ型(PIV-Ⅰ)、副流感病毒Ⅱ型(PIV-Ⅱ)、副流感病毒Ⅲ型(PIV-Ⅲ)7种呼吸道病毒进行检测以了解其混合感染情况,并结合病例临床资料对其流行病学特征进行分析。 结果 4 232例NPA标本中,HBoV检出率为1.23%(52/4 232)。1～<3岁儿童HBoV阳性检出率最高,为2.81%(24/854)。HBoV在春夏秋冬各季节的检出率分别为3.05%、0.60%、0.52%、0.24%, 差异均有统计学意义(χ²=36.784,P=0.002),其中以春季检出率最高。HBoV与其他病毒混合感染率为69.23%(36/52)。以支气管肺炎患儿HBoV阳性检出率(51.92%)最高。 结论 2014-2015年HBoV为武汉地区儿童发热呼吸道症候群的主要病原之一,1～<3岁儿童检出率最高,与其他病毒有较高的混合感染率,且春季流行。     

2011-2018年北京市通州区儿童急性呼吸道感染九种病毒性病原体监测研究

目的 了解北京市通州区儿童急性呼吸道感染病毒性病原体感染情况及流行病学特征,为疾病监测和防控策略的制定提供依据。方法 选取2011年1月-2018年6月首都医科大学附属北京潞河医院就诊的儿童急性呼吸道感染病例,使用多重实时荧光PCR检测九种呼吸道病毒性病原体,分析各呼吸道病毒性病原体感染情况。结果 544例急性呼吸道感染儿童中,呼吸道病毒性病原体检测阳性者174例,阳性率为32.0%。其中鼻病毒和流感病毒的阳性率分别为9.4%和8.6%,高于其他病原体。各季节呼吸道病原体阳性率比较,差异无统计学意义;不同季节病原体构成比较,差异有统计学意义。病原体阳性患儿流涕发生率高于阴性患儿（P=0.015）;急性上呼吸道感染患儿中呼吸道病原体阳性率高于肺炎患儿中的阳性率（43.9% vs 22.3%,P<0.001）。相比较肺炎,流感病毒、鼻病毒、偏肺病毒、冠状病毒和肠道病毒更易在急性上呼吸道感染中检出。结论 通州区儿童急性呼吸道感染主要由鼻病毒和流感病毒两种呼吸道病原体所致,呼吸道病毒更易引起急性上呼吸道感染。     

武汉市呼吸道感染儿童博卡病毒的检测及临床分析

目的 分析武汉市呼吸道感染儿童博卡病毒(HBoV)感染的临床特点,为临床治疗提供理论依据。方法 收集2014年12月1日-2015年12月1日期间在本院儿科住院的365例急性呼吸道感染患儿为研究对象,在疾病急性期用一次性吸痰管采集患儿鼻咽分泌物(NPA)标本液2 ml。先进行7种常见呼吸道病毒的筛查,然后采用PCR方法检测HBoV 基因,对阳性标本进行序列验证及分析。结果 1)对365份标本进行免疫荧光检测,结果显示呼吸道合胞病毒(RSV)的阳性检出率最高18.90%(69/365),其他依次为腺病毒(ADV)阳性15例(4.11%),副流感病毒Ⅲ(PIV-Ⅲ)阳性8例(2.20%),甲型流感病毒(IFV-A)阳性5例(1.37%),乙型流感病毒(IFV-B)阳性3例(0.82%),副流感病毒Ⅱ型(PIV-Ⅱ)阳性2例(0.55%)。2)365例标本中,32例(8.77%)出现HBoV 的PCR扩增产物,均为单一条带,大小约为354 bp,与预期的扩增片段大小相符合,显示为HBoV阳性。32例阳性标本进行测序后经Blast分析,HBoV1型28例(7.67%),HBoV2型3例(0.82%),HBoV3型1例(0.27%)。3)32例HBoV阳性患儿中,其中男19例,女13例,男女比例为1.47∶1;年龄3~13个月,平均6.87个月。结论 HBoV 感染是引起武汉市小儿呼吸道感染的重要病毒之一,主要以HBoV1型感染为主,而且多为混合感染。应加强对婴幼儿的预防保健工作,以便进行合理用药、早期治疗。     

Clinical Assessment and Improved Diagnosis of Bocavirus-induced Wheezing in Children, Finland

Human bocavirus (HBoV) is a widespread respiratory virus. To improve diagnostic methods, we conducted immunoglobulin (Ig) G and IgM enzyme immunoassays with recombinant virus–like particles of HBoV as antigen. Acute-phase and follow-up serum samples from 258 wheezing children and single serum samples from 115 healthy adults in Finland were examined. Our assays had a sensitivity of 97% and a specificity of 99.5%. Of adults, 96% had immunity; none had an acute infection. Of 48 children with serologically diagnosed acute HBoV infections, 45 were viremic and 35 had virus in nasopharyngeal aspirates (NPAs). Of 39 HBoV NPA PCR–positive children co-infected with another virus, 64% had a serologically verified HBoV infection. HBoV caused illness of longer duration than rhinovirus and of equal severity to that of respiratory syncytial virus. Among children with bronchiolitis, >25% had acute HBoV infections. Accurate HBoV diagnosis requires serologic analysis or PCR of serum; PCR of NPAs alone is insufficient.     

应用多重PCR技术快速检测临床标本中常见呼吸道病毒

[目的]应用多重PCR技术快速检测流感样病例和不明原因肺炎病例呼吸道标本中的常见呼吸道病毒,为急性病毒性呼吸道感染的监测和应急处理提供快速诊断手段。[方法]提取标本中病毒核酸RNA/DNA,反转录合成cDNA,多重PCR扩增目的基因,电泳检测扩增产物。[结果]从30份呼吸道标本中检出甲型流感病毒、乙型流感病毒、副流感病毒3型和呼吸道合胞病毒A型。[结论]本研究初步验证了该多重PCR技术能用于快速检测流感样病例和不明原因肺炎病例呼吸道标本中的常见呼吸道病毒。     

Association of human bocavirus 1 infection with respiratory disease in childhood follow-up study, Finland

Human bocavirus 1 (HBoV1) DNA is frequently detected in the upper airways of young children with respiratory symptoms. Because of its persistence and frequent co-detection with other viruses, however, its etiologic role has remained controversial. During 2009-2011, using HBoV1 IgM, IgG, and IgG-avidity enzyme immunoassays and quantitative PCR, we examined 1,952 serum samples collected consecutively at 3- to 6-month intervals from 109 constitutionally healthy children from infancy to early adolescence. Primary HBoV1 infection, as indicated by seroconversion, appeared in 102 (94%) of 109 children at a mean age of 2.3 years; the remaining 7 children were IgG antibody positive from birth. Subsequent secondary infections or IgG antibody increases were evident in 38 children and IgG reversions in 10. Comparison of the seroconversion interval with the next sampling interval for clinical events indicated that HBoV1 primary infection, but not secondary immune response, was significantly associated with acute otitis media and respiratory illness.     

Prevalence analysis of different human bocavirus genotypes in pediatric patients revealed intra-genotype recombination

BackgroundHuman bocavirus (HBoV) genotypes 1–4 have been detected worldwide in respiratory samples and stool samples, and are increasingly associated with respiratory and intestinal infections of previously unknown etiology in young children. Several studies revealed evidence of extensive recombination among HBoV genotypes at the NP1 and VP1 gene boundary region. This study explored the prevalence of HBoV genotypes in pediatric patients in Beijing, and studied their phylogeny.MethodsA total of 4941 respiratory specimens and 1121 fecal specimens were collected from pediatric patients with respiratory infections from January 2006 to December 2013, or with acute diarrhea from October 2010 to December 2012. Conventional PCR was used to detect HBoV1–4 within these samples. Gene fragments at the NP1 and VP1 gene boundary were amplified from HBoV-positive specimens, sequenced, and their phylogenetic inferences constructed using MEGA 6.0 software. Recombination events were identified with SimPlot software.ResultsHuman bocavirus 1, 2, and 3 were detected in 9 (0.80%), 15 (1.33%), and 1 (0.08%) of 1121 stool samples, respectively. However, only HBoV1 (82, 1.65%) was detected in respiratory specimens. Phylogenetic analysis of gene fragments at the HBoV NP1 and VP1 gene boundary indicated that HBoV1 sequences obtained from fecal or respiratory specimens across 8 years were highly conserved (99–100%), while 15 HBoV2 sequences collected across 2 years in Beijing were more diverse with up to 4.40% variation. Of the 15 HBoV2 sequences, 14 clustered into a new lineage divergent from other HBoV2 sequences in GenBank. Five HBoV2 genomic sequences were analyzed for recombination, revealing intra-genotype recombination between HBoV2A and HBoV2B.ConclusionsMore HBoV1 were detected in children with respiratory tract diseases, and HBoV2 in patients with acute diarrhea. Phylogenetic analysis revealed a new cluster of HBoV2 was prevalent in China, which may be the result of intra-genotype recombination between HBoV2A and HBoV2B.     

Novel Human Bocavirus in Children with Acute Respiratory Tract Infection

Human bocavirus (HBoV) and HBoV2, two human bocavirus species, were found in 18 and 10 of 235 nasopharyngeal aspirates, respectively, from children hospitalized with acute respiratory tract infection. Our results suggest that, like HBoV, HBoV2 is distributed worldwide and may be associated with respiratory and enteric diseases.     

Circulating of human bocavirus 1, 2, 3, and 4 in pediatric patients with acute gastroenteritis in Thailand

Human bocavirus (HBoV) is a newly identified human parvovirus that associated with respiratory and gastrointestinal diseases. Epidemiological surveillance of HBoV was conducted on fecal specimens collected from hospitalized children with diarrhea in Chiang Mai, Thailand in 2011. Among a total of 222 fecal specimens tested, 17 (7.7%) were positive for HBoV by PCR. Of the 17 HBoV positive samples, double- or triple-infections together with other enteric viruses were found in 10 (58.8%) pediatric patients, while monoinfection with HBoV alone was detected in seven (41.2%) cases. Mixed infection among HBoV with norovirus GII was frequently observed in this population. The partial VP1 nucleotide sequences of all 17 HBoV strains demonstrated that all four species of HBoV were found in the specimens tested. Eleven strains were HBoV1. Other three strains showed the sequence identity with HBoV2, which were most closely related to the HBoV2A. In addition, other two HBoV strains showed the highest level of nucleotide sequence identity with the HBoV3. It was surprisingly to observe that one Thai HBoV strain showed a unique characteristic similar to the HBoV4, a rare species of HBoV found in acute gastroenteritis patients. In summary, this study presents the genetic background information of HBoV circulated in acute gastroenteritis children in Chiang Mai, Thailand and it was clearly demonstrated that HBoVs circulated in this area were genetically diverse as all four species of HBoVs (HBoV1-4) were detected in the fecal specimens collected from pediatric patients admitted to the hospitals in this area.