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。     

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.     

2010～2011年乌鲁木齐地区住院呼吸道感染患儿人博卡病毒检测与分析

目的 了解新疆乌鲁木齐地区住院呼吸道感染患儿中人博卡病毒(human bocavirus,HBoV)的检出情况.方法 收集新疆维吾尔自治区人民医院2010年1月至2011年12月住院呼吸道感染患儿鼻咽抽吸物标本525例,用巢氏PCR扩增人HBoV NS1片段检测HBoV 1 ～4型.结果 在525份标本中,HBoV总阳性检出率为8.38％ (44/525),其中HBoV 1型6例,HBoV 2型38例,未检出HBoV 3、4型.各民族患儿之间HBoV阳性检出率统计分析发现,维族、回族患几分别高于汉族患儿,差异有统计学意义(P＜0.05).HBoV 1型与参考株核苷酸序列相似度为97.7％ ～ 99.4％,HBoV2型与参考株核苷酸序列相似度为93.6～ 100％.HBoV的检出时间主要以冬春季为主,HBoV感染在年龄及性别方面差异均无统计学意义.结论 2010～ 2011年乌鲁木齐地区呼吸道感染的住院患儿存在HBoV 1、2型的流行,以HBoV 2型为主,维族、回族患儿HBoV的检出率分别高于汉族患儿.     

乌鲁木齐地区腹泻患儿中人博卡病毒感染分子流行病学调查

目的 了解新疆乌鲁木齐地区腹泻患儿中人博卡病毒1～4型(HBoV1 ～4)感染的分子流行情况.方法 收集新疆维吾尔自治区人民医院2011年1月至12月住院及门诊腹泻患儿粪便标本315例,用巢氏PCR扩增入博卡病毒(HBoV) NS1片段(518 bp),检测HBoV1～4型.结果 315份标本中,HBoV总阳性检出率为8.57％ (27/315),其中HBoV1、2、3、4型分别为2例、22例、3例、0例.除XJ1378外,其他26例HBoV均与参考株的核苷酸同源性达到98％ ～ 100％,但其中3例HBoV3型与大猩猩GBoV1型(HM145750.1)核苷酸同源性为92％,且系统进化显示HBoV3型NS1片段更接近于HBoV1型.HBoV感染呈全年散发,并无明显季节性.在性别、年龄及民族间均无差异.结论 本地区腹泻患儿中HBoV1 ～3型均有流行,且以HBoV2型为主要流行株.     

北京地区婴幼儿急性呼吸道感染与新近报道的人细小病毒相关性的初步研究

目的了解北京地区婴幼儿急性呼吸道感染与新近报道的人细小病毒（human bocavirus,HBoV）的关系.方法选择2003年11月至2004年2月收集的经间接免疫荧光和/或病毒分离排除了常见呼吸道病毒感染的急性呼吸道感染患儿标本319例,应用针对HBoV的NP1基因的PCR引物进行HBoV基因片段检测,随机选取HBoV基因检测阳性标本中的5例,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.结果319例标本中HBoV基因检测阳性的为13例,其阳性检出率占本组检测标本的4.1%;HBoV阳性检出率在本组的毛细支气管炎患儿中最高,达10.9%（5/46）,其次为支气管炎患儿（6.3%,2/32）;HBoV检测阳性标本的患儿年龄主要分布于5个月～5岁,尤其是＜1岁的患儿;其中6～7个月的患儿中HBoV检测阳性的占50%（2/4）,9～10个月的患儿中阳性的占25%（2/8）,其次为5～6个月的患儿（18.2%）、11个月～1岁的患儿（14.3%）及8～9个月的患儿（12.5%）;在＜5个月的总计103例患儿中以及＞5岁的28例患儿中均未检测到HBoV阳性标本;基因序列分析表明,本研究中5例北京的HBoV之间基因序列的同源性在99.7%～100%之间;与st1、st2株的同源性为99.2%～99.4%.结论本研究结果提示北京地区部分儿科患者的急性呼吸道感染与HBoV有关,且HBoV感染在低年龄组儿童中更为常见.     

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。     

2007—2011年长沙地区急性下呼吸道感染住院儿童人偏肺病毒的流行状况

目的了解人偏肺病毒（hMPV）在长沙地区急性下呼吸道感染住院患儿中的流行病学特点。方法收集2007年9月至2011年2月因急性下呼吸道感染在湖南省人民医院儿科医学中心住院儿童的鼻咽抽吸物（nasopharyngealaspirates,NPA）样本2613份,采用逆转录聚合酶链反应法（RT—PCR）检测hMPVM基因,将阳性PCR扩增产物测序并与GenBank中已知的hMPV参考株进行比对、分析。结果2613份标本中hMPV阳性检出数135例,检出率为5．2％,男女之间的检出率比较有统计学差异（x2=8．007,P=0．003）,hMPV阳性检出患儿的年龄以1岁以内多见（63．2％）。hMPV阳性检出率在春季呈现高峰,从检出季节分布显示A2b型主要在冬春季节流行,而B2型主要在春夏季流行。135例hMPV长沙株分为A型和B型两个主要的基因型,其中A2b亚型在2007--2008年为优势流行型别,2009--2010年A2b和B2型共同流行,B2亚型在2011年呈优势流行型别。135例hMPV检出阳性患儿中有66例（48．9％）存在混合病毒感染,其中与HBoV混合检出率最高。结论长沙地区部分儿童的急性下呼吸道感染与hMPV有关,且阳性检出患儿年龄主要集中在1岁以下,男多于女,主要流行季节在春季,A2b型和B2型优势基因型在长沙地区共同流行,与其他病毒混合检出率较高。     

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，     

北京地区呼吸道感染住院儿童中人博卡病毒流行病学和基因进化分析

目的:分析北京地区呼吸道感染住院儿童人博卡病毒1型(human bocavirus 1,HBoV1)的流行病学特征,阐明遗传进化特点。方法:采用real-time PCR方法对2017—2019年北京友谊医院呼吸道感染住院患儿的2 848份鼻咽抽吸物样本进行HBoV1核酸检测,结合临床信息进行流行病学分析;利用巢氏PC...     

南京地区儿童人偏肺病毒感染的流行病学及临床特征分析

目的了解南京地区儿童人偏肺病毒（hMPV）感染的流行病学特点及临床特征。方法收集2009年8月至2010年7月南京医科大学附属南京儿童医院住院及门诊呼吸道感染患儿的鼻咽抽吸物（NPA）及咽拭子（NPS）共642例,采用逆转录聚合酶链反应法（RT—PCR）检测hMPVM基因,将阳性PCR扩增产物进行测序、同源性和进化分析。结果642例标本中共检出hMPV阳性扩增产物35份,检出率为5．5％。系统进化分析显示南京地区hMPVB1型占51．4％,A2b型占31．4％。hMPV的发病高峰在4月份。其致呼吸道感染以1岁以内多见（71．4％）。35例hMPV感染患儿中有15例（42．8％）存在混合感染,其中与HRV的混合感染检出率最高。临床诊断以肺炎（17例,48．6％）最为常见。结论人偏肺病毒是南京地区儿童急性呼吸道感染的重要病原之一,该年度其优势流行型别为B1型,南京地区A、B两型hMPV感染患儿临床特征无明显差异。     

Human bocavirus infection in young children with acute respiratory tract infection in Lanzhou,China

Human bocavirus (HBoV) is a recognized human parvovirus associated with acute respiratory tract infection. However, HBoV has yet to be established as a causative agent of respiratory disease. In this study, the epidemiological and virological characteristics of HBoV infection were studied in children with acute respiratory tract infection in China. In total, 406 children younger than 14 years of age with acute respiratory tract infection were included in this prospective 1‐year study. HBoV was detected in 29 (7.1%) of the 406 children. No clear seasonal fluctuation was observed in infection rates of HBoV. Of the 29 children infected with HBoV, 16 (55.2%) were coinfected with other respiratory viruses, most commonly respiratory syncytial virus (RSV). Viral coinfection with HBoV did not affect the severity of the respiratory disease (P = 0.291). The number of HBoV genome copies ranged from 5.80 × 10² to 9.72 × 10⁸ copies/ml in nasopharyngeal aspirates among HBoV‐positive specimens by real‐time PCR, and neither coinfection nor the severity of disease correlated with the viral load (P = 0.148, P = 0.354, respectively). The most common clinical features were cough and acute upper respiratory infection, and acute bronchopneumonia. Additionally, the NP‐1 gene of HBoV showed minimal sequence variation. These data suggest that HBoV is frequent in young children with acute respiratory tract infection in Lanzhou, China, and RSV is the most common coinfecting virus. There was no apparent association between the viral load of HBoV and coinfection or disease severity. The NP‐1 gene was highly conserved in HBoV. J. Med. Virol. 82:282–288, 2010. © 2009 Wiley‐Liss, Inc.     

Newly identified respiratory viruses associated with acute lower respiratory tract infections in children in Lanzou,China, from 2006 to 2009

Nasopharyngeal aspirates were collected from 813 children <14 years old with acute lower respiratory tract infections in Lanzhou, China, from December 2006 to November 2009. PCR or RT-PCR was used to screen for the presence of 10 respiratory viruses. Viral agents were identified in 73.92% (601/813) of specimens, including RSV in 40.71%, hMPV in 6.15%, IFVA in 7.13%, IFVB in 0.98%, PIV1-3 in 7.87%, HCoV-HKU1 in 2.21%, HCoV-NL63 in 3.81%, HRV in 19.93%, AdV in 7.50% and HBoV in 11.56%. Two or more viruses were detected in 34.44% (280/813) of cases. The newly identified respiratory viruses, HBoV, hMPV, HCoV-HKU1 and HCoV-NL63, accounted for 22.01% of the detected viral pathogens. RSV and HRV were frequently detected in patients with bronchiolitis, and hMPV was frequently associated with pneumonia. HCoV-NL63 was found to be one of the causative agents of acute respiratory wheezing in young children. No seasonal variation was found in the incidence of detection of HCoV-HKU1, HCoV-NL63 or HBoV. This 3-year study demonstrated that viral pathogens play an important role in children with ALRTIs, and more attention should be paid to these newly identified viral agents.     

Human bocavirus in children with acute respiratory infections in Vietnam

Acute respiratory infections are the major cause of morbidity and mortality globally. Human bocavirus (HBoV), a novel virus, is recognized to increasingly associate with previously unknown etiology respiratory infections in young children. In this study, the epidemiological, clinical, and molecular characteristics of HBoV infections were described in hospitalized Vietnamese pediatric patients. From April 2010 to May 2011, 1,082 nasopharyngeal swab samples were obtained from patients with acute respiratory infections at the Children's Hospital 2, Ho Chi Minh City, Vietnam. Samples were screened for HBoV by PCR and further molecularly characterized by sequencing. HBoV was found in 78 (7.2%) children. Co‐infection with other viruses was observed in 66.7% of patients infected with HBoV. Children 12–24 months old were the most affected age group. Infections with HBoV were found year‐round, though most cases occurred in the dry season (December–April). HBoV was possible to cause severe diseases as determined by higher rates of hypoxia, pneumonia, and longer hospitalization duration in patients with HBoV infection than in those without (P‐value <0.05). Co‐infection with HBoV did not affect the disease severity. The phylogenetic analysis of partial VP1 gene showed minor variations and all HBoV sequences belonged to species 1 (HBoV1). In conclusion, HBoV1 was circulating in Vietnam and detected frequently in young children during dry season. Acute respiratory infections caused by HBoV1 were severe enough for hospitalization, which implied that HBoV1 may have an important role in acute respiratory infections among children. J. Med. Virol. 86:988–994, 2014. © 2013 Wiley Periodicals, Inc.

     

Human Bocavirus quantitative DNA detection in French children hospitalized for acute bronchiolitis

BACKGROUND: Human Bocavirus (HBoV) is a newly discovered parvovirus whose role as a causative agent of respiratory disease remains unclear. STUDY DESIGN: We investigated the presence of HBoV by quantitative PCR in the nasopharyngeal samples of 192 French children consecutively hospitalized for acute bronchiolitis. Other common respiratory viruses were detected using immunofluorescence assays, cell culture detection, or RT-PCR assays. RESULTS: HBoV was detected in 24 (12.5%) of 192 study children. In 14/192 cases (7%) HBoV was the sole isolate and in 10/192 (5%) it was part of a mixed viral infection. HBoV was the third most common pathogen detected after respiratory syncytial virus (45/192; 23%) and rhinovirus (24/192; 12%). It occurred more often in infants aged 1-12 months (P=0.002). Median levels of HBoV DNA genome in respiratory samples were significantly higher in patients with single HBoV infection than in patients with mixed respiratory viral infection with HBoV (4x10(8)copies/ml vs. 2x10(3)copies/ml, P<0.001). CONCLUSIONS: Our data suggest that HBoV at a high viral load could be an etiologic agent of respiratory tract disease, whereas the exact role of HBoV at a low viral load, as etiological cause or as pathophysiological co-factor of respiratory diseases, remains to be determined.     

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.     

Occurrence of human bocaviruses and parvovirus 4 in solid tissues

Human bocaviruses 1-4 (HBoV1-4) and parvovirus 4 (PARV4) are recently discovered human parvoviruses. HBoV1 is associated with respiratory infections of young children, while HBoV2-4 are enteric viruses. The clinical manifestations of PARV4 remain unknown. The objective of this study was to determine whether the DNAs of HBoV1-4 and PARV4 persist in human tissues long after primary infection. Biopsies of tonsillar tissue, skin, and synovia were examined for HBoV1-4 DNA and PARV4 DNA by PCR. Serum samples from the tissue donors were assayed for HBoV1 and PARV4 IgG and IgM antibodies. To obtain species-specific seroprevalences for HBoV1 and for HBoV2/3 combined, the sera were analyzed after virus-like particle (VLP) competition. While HBoV1 DNA was detected exclusively in the tonsillar tissues of 16/438 individuals (3.7%), all of them ≤8 years of age. HBoV2-4 and PARV4 DNAs were absent from all tissue types. HBoV1 IgG seroprevalence was 94.9%. No subject had HBoV1 or PARV4 IgM, nor did they have PARV4 IgG. The results indicate that HBoV1 DNA occurred in a small proportion of tonsils of young children after recent primary HBoV1 infection, but did not persist long in the other tissue types studied, unlike parvovirus B19 DNA. The results obtained by the PARV4 assays are in line with previous results on PARV4 epidemiology.     

Human bocavirus infections in hospitalized Greek children

Introduction

The epidemiology of human bocavirus (HBoV) infections has not been described in Greece, a south-eastern European country. To define the epidemiological profile and the clinical characteristics associated with HBoV infection in a population of children hospitalized with respiratory tract infection.

Material and methods

During a one-year period throat swab samples were collected from 370 previously healthy children, aged 14 days to 13 years, admitted to two different paediatric wards because of respiratory tract infection. Samples were tested for HBoV by PCR amplifying a part of the NS1 gene.

Results

Human bocavirus was detected in 12 children (3.2%). Four of the 12 cases were co-infections, 3 of them with influenza A and 1 with coronavirus OC43. Cases were observed only during the cold months. The mean age of children was 1.8 years (range 2 months to 4 years). The most common symptoms were fever, cough and various degrees of respiratory distress. All children were clinically diagnosed as having lower respiratory tract infections, mainly pneumonia and acute laryngotracheobronchitis, and recovered uneventfully.

Conclusions

HBoV infections occur in Greece mostly among very young children. They accounted for 3.2% of children hospitalized with acute respiratory disease. Cases were observed only in late autumn to early spring.     

Detection of human bocaviruses 1 to 4 from nasopharyngeal swab samples collected from patients with respiratory tract infections

Human bocaviruses (HBoV) 1, 2, 3, and 4 (HBoV1-4) were detected in 132 (15.5%), 5 (0.6%), 3 (0.4%), and 5 (0.6%) of 850 nasopharyngeal swab samples collected from children with respiratory tract infections, respectively. Out of the 145 HBoV1-4-positive samples, 62 (42.8%) were codetected with other respiratory viruses.     

Correlation between bocavirus infection and humoral response,and co-infection with other respiratory viruses in children with acute respiratory infection

BackgroundHuman bocavirus (HBoV), a recently discovered virus, is prevalent among children with respiratory tract infection throughout the world. Co-infection was frequently found in HBoV-positive patients. Thus, whether HBoV is responsible for the respiratory disease is still arguable.ObjectivesA comprehensive study was carried out to integrate clinical and virological prevalence in HBoV-positive outpatient children, and to determine genetic and serologic characteristics of HBoV in Shanghai, China.Study designNasal/throat swabs and sera were obtained over a 2-year period from 817 children with respiratory tract infection to examine the presence of HBoV and its co-infection. The seroepidemiology of HBoV was studied by ELISA and Western blot against the capsid protein VP2-based fragment. Persistence of HBoV was also analyzed in 12 pairs of return-visit cases.ResultsHBoV was identified in 96 samples (11.8%). The co-infection rate with other respiratory viruses was 51%. IgM was detected in 55.7% of HBoV RT-PCR-positive patients, and in 72.7% of those who had high viral genome load. In addition, persistent viral DNA positivity was detected in 10 of 12 HBoV-positive cases tested, an average of 14 days later, and one child was still HBoV-positive after 31 days.ConclusionHBoV was found frequently in children with respiratory tract symptoms associated with other respiratory viruses, and persisted in the respiratory tract and in serum and urine. The presence of IgM was significantly more prevalent in viremic patients and those diagnosed with high load of HBoV DNA in nasal/throat swabs.