重症急性呼吸道感染儿童中人博卡病毒的分型检测与基因进化分析

目的 了解重症急性呼吸道感染住院儿童中人博卡病毒（HBoV）的感染状况,流行病学特征及其进化特征.方法 采用巢式PCR的方法,对来自北京儿童医院重症急性呼吸道感染住院儿童的259份鼻咽抽吸物,进行人博卡病毒（HBoV）分型检测与测序,同时进行了合并感染检测、流行病学、临床特点及基因多态性分析.结果 共检出56份人博卡病毒感染阳性标本,阳性率为21.6％,[95％ CI（16.0％～27.3％）,P＜0.0001],其中2岁以下儿童感染率较高.与其他呼吸道常见病毒的合并感染率为94.6％.HBoV阳性产物测序分析发现,人博卡病毒1型占96.4％ （54/56）,2、3型各1份.HBoV阳性株分型区（VP1/VP2）序列变异不明显.结论 人博卡病毒是儿童急性呼吸道感染常见的病原体,以I型最为常见,分型区（VP1/VP2）序列较保守.HBoV在重症急性呼吸道感染儿童中是否起到真正的致病作用还需进一步的研究.     

儿童急性呼吸道博卡病毒感染的病毒载量与临床特征相关性研究

目的 研究急性呼吸道感染患儿人博卡病毒（ human bocavirus,H BoY）病毒载量与临床特征的相关性。方法 对2009年l1月至2010年12月间956例呼吸道感染的患儿及251例健康对照组儿童鼻咽部抽吸物、咽拭子采用PCR法进行HBoV检测,进而对阳性样本进行实时荧光定量PCR法测定博卡病毒DNA载量,并结合患儿的临床检查进行综合分析。结果 实验组与对照组HBoV阳性率存在显著差异,下呼吸道感染病例HBoV的病毒载量水平与上呼吸道感染病例及对照组儿童差异均有统计学意义,上呼吸道感染病例与对照组儿童病毒载量无统计学意义,重症下呼吸道感染患儿与普通下呼吸道感染患儿HBoV的病毒载量无统计学差异,HBoV混合感染与独立感染患儿病毒载量亦无统计学差异。结论 博卡病毒常年均可引起发病,是儿童呼吸道感染的重要病原体之一,但可能不是儿童急性呼吸道感染的唯一因素。HBoV病毒载量并不能独立反映临床疾病感染的严重程度。     

我国急性呼吸道感染患儿中检测到KI和WU多瘤病毒

目的 了解多瘤病毒WU和KI在我国儿童急性呼吸道疾病中的感染情况.方法 采用PCR扩增的方法对2006年11月至2007年10月收集的急性呼吸道感染患儿的318份鼻咽抽吸物(NPA)标本进行了多瘤病毒WU和KI基因检测.结果 318份标本共检测出14份病毒核酸阳性标本,其中WUV 7份(2.2%),KIPyV 7份(2.2%).该14例基因检测阳性患儿临床均有上呼吸道感染或下呼吸道感染症状.结论 WUV和KIPyV可能也是儿童急性呼吸道感染中较为重要的一个病原,且与儿童上呼吸道感染和下呼吸道感染存在相关性.     

兰州地区呼吸道感染患儿中人类博卡病毒1～3型检测与临床研究

目的了解兰州地区急性呼吸道感染患儿中人博卡病毒1—3型（HBoV1～3）感染的临床及分子流行病学特征。方法收集兰州大学第一医院2009年12月至2010年11月急性呼吸道感染患儿鼻咽分泌物及咽拭子标本524份,用：巢氏PCR扩增人博卡病毒（HBoV）NS1片段,检测HBoV1～3;同时PCR检测常见呼吸道病毒。结果524份标本中检出HBoV43例,检出率为8．2％,仅次于鼻病毒、呼吸道合胞病毒、副流感病毒3型;混合感染率为69．8％。其中人博卡病毒1型（HBoV1）在下呼吸道感染中检出率显著高于上呼吸道感染的检出率;2例人博卡病毒2型（HBoV2）患儿都出现胃肠道症状,与标准株GU048662．1的核苷酸同源性分别为99％和100％;1例人博卡病毒3型（HBoV3）与标准株HM132056．1的核苷酸同源性为99％。结论本地区儿童急性呼吸道感染中博卡病毒感染以HBoV1为主,首次检出HBoV3;人博卡病毒与其他病毒有较高的合并感染。人博卡病毒是本地区儿童急性呼吸道感染的重要病原之一。     

上海地区人博卡病毒在儿童急性呼吸道感染的研究CSCD

目的了解上海地区人博卡病毒（HBoV）在儿童急性呼吸道感染中的流行情况和临床特点。方法上海地区在2012年1月至2012年12月共收集271例急性呼吸道感染患儿的鼻咽抽吸物,采用巢式聚合酶链反应（Nested PCR）方法检测人博卡病毒NS1基因、并经测序确认,对所获得的基因序列进行同源性和进化分析,博卡病毒阳性样本同时检测鼻病毒、呼吸道合胞病毒、腺病毒等8种呼吸道相关病毒。结果271例标本中共检出HBoV阳性31例,检出率11.4%;21例存在混合感染,全年均有检出,阳性患儿中位年龄17个月（4个月-4岁）,诊断包括上呼吸道及下呼吸道感染,临床表现包括发热、阵发性咳嗽、咳痰、腹泻、呕吐、咽充血、湿罗音等,无死亡病例,门诊患者检出率明显高于住院患者;序列分析表明其中29例为HBoV1、2例为HBoV2、与参考株的核苷酸同源性为99%-100%,氨基酸同源性96%-100%。结论HBoV1是上海地区急性呼吸道感染患儿中的重要病原,HBoV2在该地区首次检出,临床症状及诊断无特异性。     

人博卡病毒研究进展

2005年，瑞典的Allander等人^[1]采用宿主DNA消除、随机PCR扩增、高通量测序和生物信息学知识相结合的方法，在呼吸道感染患儿的鼻咽抽吸物中发现了一种新的细小病毒——人博卡病毒（hu．man bocavirus，HBoV）。随后，世界上许多国家陆续报道了在呼吸道感染患儿中检测出HBoV，     

长沙地区呼吸道感染儿童Saffold心肌炎病毒的调查

目的了解长沙地区Saffold病毒（以下简称为SAFV）在儿童呼吸道感染中的流行情况,探讨其与儿童呼吸道感染的相关性。方法选取湖南省人民医院儿科医学中心2007年11月至2008年10月间643名因呼吸道感染住院儿童的鼻咽抽吸物。采用实时荧光定量聚合酶链式反应（Realtime．PCR）方法扩增SAFV的5UTR的基因片段,并且统计分析临床资料。结果643份样本中共检测出SAFV阳性67份,阳性检出率为10．42％（67／643）,5岁以上未检测出该病毒。31例患迁延性肺炎和慢性肺炎的患儿标本检出SAFV8例（25．81％）,差异有统计学意义。结论本研究表明长沙地区下呼吸道感染住院儿童存在SAFV感染;SAFV可能与下呼吸道感染及病程迁延相关。     

长沙地区住院儿童冠状病毒HKU1的感染状况及临床特征

目的了解冠状病毒HKU1在长沙地区急性下呼吸道感染住院儿童中的感染状况及临床特征。方法采集2007年9月至2008年8月长沙地区因急性下呼吸道感染住院儿童鼻咽抽吸物（nasopharyngeal aspirates,NPA）样本1165份,用RT-PCR方法检测冠状病毒HKU1 pol基因,测序分析阳性扩增产物,统计分析临床资料。结果1165份NPA样本中,检测出12份冠状病毒HKU1,阳性率为1．03％（12／1165）,患儿年龄最小8天,最大3岁,临床诊断多见支气管肺炎（83．33％）,所测核苷酸序列与冠状病毒HKU1参考株同源性为98．18％～100％。结论冠状病毒HKU1阳性患儿年龄均在3岁及3岁以下;性别无差异;冬春季检出率高;单一基因型在长沙地区流行;HKU1也是我国儿童急性下呼吸道感染中的一种病原。     

人细小病毒——博卡病毒的研究进展

2005年8月22日瑞典学者Tobias Allander[1]在世界首次报道从呼吸道感染患者病理标本中分离出一种新型的人细小病毒,并命名为人博卡病毒(Human Bocavirus,HSoV).随后在全球相继有该型病毒感染的报道,初步判断人博卡病毒是引起急性呼吸道感染的一种新型重要病原.本文将对该型病毒的临床感染情况和分子生物学研究方面作一综述.     

兰州地区急性呼吸道感染患儿冠状病毒NL63的检测与临床研究

目的 探讨兰州地区冠状病毒NL63(HCoV-NL63)在急性呼吸道感染患儿中的流行现状及临床特点.方法 收集2006年11月至2009年10月兰州大学第一附属医院急性呼吸道感染(ARTIs)患儿1169例鼻咽分泌物,应用RT-PCR方法检测HCoV-NL63以及其余7种常见呼吸道病毒:鼻病毒(HRV),呼吸道合胞病毒(RSV),偏肺病毒(hMPV),流感病毒(IFVA,IFVB)副流感病毒1-3(HPIV1-3)及PCR方法检测腺病毒(ADV),博卡病毒(HBoV).结果 检测出HCoV-NL63阳性标本35例,检出率2.99％,2007年8、9月,2009年7、8月检测阳性标本阳性率较高,分别为23.53％、17.65％,50％、33.33％.2007年12月至2009年2月未检出HCoV-NL63阳性标本.25(25/35)例混合其他病毒感染,混合感染率为71.43％,最常见的混合感染病毒是HRV.3岁及以下和3岁以上HCoV-NL63感染组感染率差异无统计学意义.HCoV-NL63阳性患儿主要的诊断是支气管肺炎和毛细支气管炎,主要的症状是发热和咳嗽.HCoV-NL63单独感染组和混合感染组除消化道症状外,在其余症状和临床诊断方面,差异均无统计学意义.结论 HCoV-NL63是兰州地区呼吸道感染患儿的重要病原,夏季是兰州地区HCoV-NL63感染高峰期,HCoV-NL63的流行存在年度差异.HCoV-NL63感染存在很高的混合感染率,混合感染并不加重HCoV-NL63感染的病情.     

Human bocavirus infection in children with acute respiratory tract infection in India

Human bocavirus (HBoV) is a new human parvovirus identified in children with respiratory tract disease. Nasopharyngeal aspirates were collected from 305 children <5 years of age with acute respiratory tract infection from April 2005 to March 2007 and screened for the presence of HBoV by two separate sets of a polymerase chain reaction (PCR) described previously. Twenty‐two (7.2%) children who had acute respiratory infection were found to be positive for HBoV by both sets of PCR. The main clinical symptoms were cough (95%), runny nose (64%), and fever (59%). In two samples, HBoV was identified together with respiratory syncytial virus in one sample and influenza A virus in another. HBoV appeared to have no seasonal distribution and is associated with both upper and lower respiratory tract disease in young children in India. J. Med. Virol. 82: 812–816, 2010. © 2010 Wiley‐Liss, Inc.     

Detection of human bocavirus in children with Kawasaki disease

Human bocavirus (HboV) is an emerging virus that has been implicated as a cause of acute upper and lower respiratory tract infection in children. As no serological assay is available, PCR was used to screen nasopharyngeal, serum or stool samples from 16 patients with Kawasaki disease for HBoV nucleic acid. HBoV was identified by PCR in five (31.2%) patients, suggesting that this emerging virus may also play a pathogenic role in some cases of Kawasaki disease.     

Seroepidemiology of human bocavirus in Hokkaido prefecture, Japan

A new human virus, provisionally named human bocavirus (HBoV), was discovered by Swedish researchers in 2005. A new immunofluorescence assay using Trichoplusia ni insect cells infected with a recombinant baculovirus expressing the VP1 protein of HBoV was developed, and the levels of immunoglobulin G antibody to the VP1 protein of HBoV in serum samples were measured. The overall seroprevalence rate of antibodies against the VP1 protein of HBoV in a Japanese population aged from 0 months to 41 years was 71.1% (145 of 204). The seropositive rate was lowest in the age group of 6 to 8 months and gradually increased with age. All of the children had been exposed to HBoV by the age of 6 years. A rise in titers of antibody against the VP1 protein of HBoV during the convalescent phase was observed for four patients with lower respiratory tract infections, and HBoV DNA was detected in nasopharyngeal swab and serum samples from all four patients. These results suggest that HBoV is a ubiquitous virus acquired early in life and that HBoV might play a role in the course of lower respiratory tract infections.     

Human bocavirus commonly involved in multiple viral airway infections.

BACKGROUND: Human bocavirus (HBoV) was recently discovered in children with acute respiratory tract infections. We have included a PCR for HBoV in a study on airway infections in children. OBJECTIVES: To study the occurrence of HBoV in Norwegian children, and to evaluate the results of a semiquantitative PCR. STUDY DESIGN: During a 4-month period in the winter season 2006/2007 we collected nasopharyngeal aspirations from children who were admitted to the Department of Pediatrics. All samples were examined for 17 agents with real-time PCR. RESULTS: HBoV was detected in 45 of 376 samples (12%). The occurrence of HBoV was stable during the study period. Multiple viral infections were present in 78% of the samples (42% double, 20% triple and 16% quadruple infections). RS-virus, enterovirus and human metapneumovirus were the most frequently codetected agents. In samples with a high load for HBoV, significantly fewer multiple infections were found than in the other samples. Eighty-eight percent of the 25 patients with HBoV recorded as either the only or the dominating virus, and 50% of the other patients, had lower respiratory tract infection. The difference was statistically significant. CONCLUSIONS: HBoV was frequently detected in nasopharyngeal aspirates from children with airway infections in Norway. Multiple viral infections were common among the HBoV-infected patients. Semiquantitative PCR results may be useful for interpretation of clinical relevance.     

Human bocavirus and other respiratory viral infections in a 2-year cohort of hospitalized children

Human bocavirus (HBoV) infection is reported worldwide and may cause severe respiratory tract infections. The aim of the present study was to assess the prevalence of HBoV, and other respiratory viral pathogens, in a 2-year retrospective study of children admitted to hospital, and to investigate whether viral loads of HBoV DNA were associated with severity of infection. Between April 2007 and March 2009, 891 respiratory samples from 760 children admitted to hospital with acute respiratory tract infection were tested for the presence of respiratory viruses by real-time PCR or direct immunofluorescence testing. HBoV DNA was detected by using internally controlled real-time quantitative PCR assay and 25 samples selected at random were sequenced. The virus detected most frequently was rhinovirus, followed by respiratory syncytial virus, HBoV, and human metapneumovirus. HBoV DNA was detected in 18.4% of children admitted to hospital. HBoV was the only viral pathogen detected in 66/164 (40.2%) of HBoV DNA-positive children and in 7.4% of all 891 samples. Ninety-seven percent (64/66) of children with an HBoV single infection were diagnosed as having lower respiratory tract infection. Median HBoV DNA viral load was significantly higher in children when HBoV was detected as a single pathogen. Higher HBoV DNA viral loads were associated with prematurity and age. HBoV seems to be an important and frequent pathogen in respiratory tract infections in children, and it is likely that the severity of illness is comparable to the severity of RSV illness.     

Pediatric hospitalization of acute respiratory tract infections with Human Bocavirus in Hong Kong.

BACKGROUND: Respiratory tract infections are a leading cause of morbidity and mortality globally. Human Bocavirus (HBoV) has recently been identified and implicated as an aetiologic agent of lower respiratory tract infection in children. OBJECTIVES: The prevalence of HBoV and clinical manifestations of children hospitalized for acute respiratory illness in Hong Kong were determined. STUDY DESIGN: 1906 non-duplicate nasopharyngeal aspirates obtained from children aged >1 month to 15 years of age hospitalized with respiratory tract infections during a 13-month period were investigated for the presence of HBoV by PCR. These children were admitted to the general pediatric wards in a teaching and tertiary Hong Kong hospital in a prospective study for surveillance of acute respiratory illness. RESULTS: Human Bocavirus was detected in 5.0% (95/1906) of nasopharyngeal aspirates by PCR. The percentage was highest in 25-36 months group (12%). Seasonal distribution was noted from September to February. Co-infection with human parainfluenza viruses and respiratory syncytial virus was present in 16% and 3% of cases, respectively. Gastrointestinal symptoms of vomiting, abdominal pain and diarrhoea were common, besides respiratory symptoms and fever. CONCLUSIONS: HBoV plays an important role in hospitalized children 相似文献   

Human Bocavirus: Passenger or Pathogen in Acute Respiratory Tract Infections?

Human bocavirus (HBoV) is a newly identified virus tentatively assigned to the family Parvoviridae, subfamily Parvovirinae, genus Bocavirus. HBoV was first described in 2005 and has since been detected in respiratory tract secretions worldwide. Herein we review the literature on HBoV and discuss the biology and potential clinical impact of this virus. Most studies have been PCR based and performed on patients with acute respiratory symptoms, from whom HBoV was detected in 2 to 19% of the samples. HBoV-positive samples have been derived mainly from infants and young children. HBoV DNA has also been detected in the blood of patients with respiratory tract infection and in fecal samples of patients with diarrhea with or without concomitant respiratory symptoms. A characteristic feature of HBoV studies is the high frequency of coinciding detections, or codetections, with other viruses. Available data nevertheless indicate a statistical association between HBoV and acute respiratory tract disease. We present a model incorporating these somewhat contradictory findings and suggest that primary HBoV infection causes respiratory tract symptoms which can be followed by prolonged low-level virus shedding in the respiratory tract. Detection of the virus in this phase will be facilitated by other infections, either simply via increased sample cell count or via reactivation of HBoV, leading to an increased detection frequency of HBoV during other virus infections. We conclude that the majority of available HBoV studies are limited by the sole use of PCR diagnostics on respiratory tract secretions, addressing virus prevalence but not disease association. The ability to detect primary infection through the development of improved diagnostic methods will be of great importance for future studies seeking to assign a role for HBoV in causing respiratory illnesses.     

Human bocavirus (HBoV)

The human bocavirus (HBoV) has been recently identified by means of molecular screening techniques in respiratory tract secretions from children with acute respiratory tract disease. This virus, which belongs to the Parvoviridae family, has been detected worldwide with a 5 to 10% prevalence among children with upper or lower respiratory tract infections, essentially during the winter period. A seroepidemiological study has shown that almost all the children have antibodies to HBoV by the age of five years, and HBoV infection seems to be rare in adults. HBoV is often detected in association with other respiratory viruses. This virus has also been detected in stools, but its role in gastroenteritis has not been yet established. Virological diagnostic of HBoV infection is based on the detection of viral DNA by PCR. Viral load determination by viral DNA quantitation in respiratory tract secretions could be a tool to differentiate between symptomatic HBoV infection and virus carriage.     

Real-time PCR for diagnosis of human bocavirus infections and phylogenetic analysis

The human bocavirus (hBoV) was first described in 2005 in respiratory tract samples. The clinical relevance of hBoV is still unclear. The aim of our study was to establish a real-time PCR assay for the detection and quantification of hBoV DNA, to apply the real-time assay for the analysis of stool and serum samples for the presence of hBoV DNA, and to perform a phylogenetic analysis of the hBoV positive samples. A total of 834 nasopharyngeal aspirates (NPA), 10 serum samples, and 31 stool samples of children with acute respiratory diseases were retrospectively tested. For phylogenetic analysis, 968 bp of the VP2 gene were sequenced from 69 hBoV-positive NPA samples. The qualitative results of the real-time hBoV PCR were in good agreement with a conventional hBoV PCR. We found that 12% of the NPA were positive for hBoV DNA. The median viral load in the NPA was 4.9 x 10(3) copies/ml (range, 2.7 x 10 degrees to 1.5 x 10(11) copies/ml). There was no difference of the hBoV load in NPA between children with or without known coinfection, but the load was significantly higher in children with bronchitis than in children with the diagnosis of febrile seizures. hBoV DNA was found in 1 of 10 serum samples and in 14 of 31 stool samples. hBoV sequence identity was >99% in the VP2 region. In conclusion, hBoV DNA can be found in NPA samples at very high titers. In addition to being found in the respiratory tract, hBoV was found in stool samples. The clinical relevance of these findings remains to be determined.