广东地区急性呼吸道感染儿童人类博卡病毒的流行状况

目的 了解广东地区急性呼吸道感染患儿人类博卡病毒(HBoV)的感染情况.方法 收集广东地区2007年6月至2008年5月期间呼吸道感染患儿的鼻咽分泌物447份,采用PCR法检测HBoV衣壳蛋白(VP)基因片段,阳性标本作核酸序列测定,并与基冈库中的已知序列进行序列比对和系统进化树分析.结果 447例呼吸道感染患儿标本中HBoV阳件率为5.1%.其中10例患儿与其他病毒混合感染,占阳性标本的43.5%.阳件患儿的主要临床诊断为喘息性肺炎、毛细支气管炎和支气管肺炎,年龄分布从42 d到6岁,主要集中在1岁以内,HBoV感染的季节分布偏向夏、秋及晚春.经序列比对和进化树分析.阳性株的VP基因片段与瑞典株ST1的核酸及氨基酸序列同源性分别为97.8%～98.8%及99.3%～100.0%.结论 HBoV是广东地区儿童下呼吸道感染的重要病原之一,且在1岁以内患儿中高发.该地区HBoV流行株的VP基因片段较为保守,但也存在导致氨基酸改变的突变株.     

一起急性腹泻疫情中检出的GI.8型诺如病毒的序列测定和分子特征分析

目的对湖州地区一起急性腹泻疫情中检出的GI.8型诺如病毒进行序列测定和分子特征分析。方法根据GI型诺如病毒保守序列自行设计5对引物,用一步法RT-PCR分段扩增GI.8型诺如病毒CHN/Huzhou/N10的全基因组序列,采用生物信息学相关软件对序列进行整理和分析。结果 CHN/Huzhou/N10全长7 740bp,编码区包括3个开放读码框架(ORF1~ORF3),分别长5 400bp(5~5 404nt)、1 632bp(5 388~7 019nt)和642bp(7 019~7 660nt)。RdRp区,VP1区和VP2区的系统进化分析显示CHN/Huzhou/N10属于GI.8基因型。衣壳蛋白区的氨基酸序列分析表明,与原型株Boxer/2001/US相比,CHN/Huzhou/N10VP1区的氨基酸序列共有16个变异位点,12个位于P2区。其中aa347T→S,aa397T→E这2个变异位点分别位于HBGA受体结合袋位点Ⅱ和Ⅲ内。结论本文获得了我国GI.8型诺如病毒CHN/Huzhou/N10的全基因组序列,相关序列信息可用于病毒的遗传进化研究、快速诊断试剂的研发以及疫苗的设计。     

2019年云南省埃可病毒18型分离株的全基因组分析

目的对2019年云南省埃可病毒18型(E-18)的分离株K88R3-19进行全基因组序列测定及分析,了解其进化和变异特性。方法利用RD、Vero和KMB17细胞分离病毒,提取病毒RNA,RT-PCR扩增全基因,并测序拼接获得全基因组序列。利用Mega 5.05、Geneious Basic 5.4.1和SimPlot 3.5.1等软件分析全基因组序列。结果 K88R3-19株全基因组序列长度为7 422 bp,编码含2 190个氨基酸的多聚蛋白。K88R3-19与E18/LJ/0601/2019全基因组核苷酸一致性和氨基酸同源性分别为97.6%和99.2%;与其他E-18毒株的核苷酸一致性和氨基酸同源性分别为79.7%～86.1%和95.8%～97.7%;与其他E18毒株的各区段核苷酸一致性和氨基酸同源性分别为72.7%～98.5%和91.9%～100%,其中与中国株VP1-VP4区段核苷酸的一致性为89.8%～98.7%。基于E18全长VP1序列的种系进化分析可将E18分为1-6个基因型,其中5基因型可再分为5-1～5-3 3个基因亚型。中国E-18流行株主要聚集在5-3基因亚型。K88R3-19株较其他E-18株在VP1氨基酸序列上有2个独特突变:第2和28位点分别由D→V和V→A。结论 K88R3-19分离株为5-3基因亚型,为2019年中国大陆流行株。     

手足口病患儿感染肠道病毒71型全基因组基因特征分析

目的从手足口病患儿粪便中分离肠道病毒71型(EV71),对其进行全基因组测序,了解秦皇岛市EV71全基因组基因特征,为手足口病的综合防治提供依据。方法从秦皇岛市各医院收集手足口病患儿粪便标本,进行EV71检测,阳性标本进行病毒分离及核酸提取,通过RT-PCR进行全基因组序列扩增,通过电泳、测序及序列拼接一系列操作分析其基因特征。结果用提取的核酸扩增出7 406bp的目的基因片段测序后与已记录的EV71相应序列进行比对完全一致,且测序峰图较好,可信度也较高;各基因片段间通过核苷酸序列拼接后得出,秦皇岛市医院EV71全基因组序列长7 406bp,编码区各基因片段的长度分别为:2A450bp、2B297bp、2C987bp、3A258bp、3B66bp、3C549bp、3D1 386bp、VPl 891bp、VP2 762bp、VP3 726bp、VP4 207bp。结论秦皇岛市EV71分离株为C4a型全基因组序列长7 406bp,可为当地手足口病的防控提供参考。     

福建省肠道病毒71型分离株（2010FJLY008）全基因组核苷酸序列分析

目的了解肠道病毒71型在福建省的遗传背景,追踪EV71流行过程中可能发生的变异。方法对2010年福建1例手足口病患儿标本中分离到的1株肠道病毒71型分离株（2010FJLY008）进行全基因组核苷酸序列测定,并对基因组序列进行同源性比较及遗传进化分析。结果2010FJLY008株全长7 403bp,核苷酸及编码氨基酸序列同源性比较,均与08年阜阳流行株Fuyang17.08 2同源性最高,整个基因组核苷酸同源性为98.2%,氨基酸同源性为99.5%;全长基因组核苷酸序列遗传进化分析及VP1区基因遗传进化分析,与Fuyang17.08 2株进化关系较为接近,同属于C4a基因亚型。结论2010年福建省EV71型病毒流行株（2010FJLY008）在病毒基因组结构、基因组核苷酸同源性比较、编码氨基酸同源性比较、全长基因组核苷酸序列遗传进化分析及VP1区基因遗传进化分析等方面,均与2008年阜阳流行株（Fuyang17.08 2）关系密切,未产生明显的抗原漂移及变异。     

猪瘟病毒cDNA片段的合成及克隆

以猪瘟病毒石门系毒株常规感染 PK15细胞48小时后,去上清,直接用异硫氰酸胍一步法提取细胞总 RNA。根据猪瘟病毒Alfort 株的基因组序列化学合成了8条引物,用3′端引物及提取的总 RNA 进行反转录合成第一链 cDNA,再以此单链 cDNA 为模板,用 PCR 法扩增猪瘟病毒 cDNA 片段,2%琼脂糖凝胶电泳鉴定,结果证明所扩增片段的长度与根据 Alfort 株基因组序列推测的长度相符,依次为693bp,629bp,346bp,285bp,120bp。用限制性内切酶分析进一步证明了所扩增片段的正确性。最后按标准方     

一株蝙蝠轮状病毒的分离和基因组分析

目的 了解云南地区蝙蝠轮状病毒的基因组特征和进化关系,并分离病毒。方法 采集84只云南地区蝙蝠,经解剖后收集肠道组织,混合为3个样本库后进行高通量测序。病毒全基因组扩增采用RT-PCR和RACE方法,使用RVA自动分型工具进行分型。采用MA104细胞分离培养病毒株。结果 高通量测序结果显示1个样本库中存在A组轮状病毒相关的序列。随后成功分离该蝙蝠轮状病毒并扩增获得全长或接近全长的11条基因组序列。基因组分型结果显示该病毒株属于G3-P[3]-I8-R3-C3-M3-A9-N3-T3-E3-H6基因型,命名为CHYNC82。序列分析表明,CHYNC82的4条基因片段(VP1、NSP2、NSP3和NSP4)与蝙蝠病毒株MYAS33和MSLH14亲缘关系近且进化树聚为一支;VP2与人09US7118株和猿猴TUCH株进化距离最近,处于同一进化树枝;VP6和NSP1与已知病毒同源性较低,但与蝙蝠和人来源轮状病毒具有共同进化起源;而剩余4条基因片段(VP7、VP4、VP3和NSP5)与人类病毒株包括CMH222株和MS2015-1-0001株具有最高同源性,且处于同一进化树枝。结论 推测CHYN...     

广东地区首例人博卡病毒的检出及鉴定

目的 建立人博卡病毒(HBoV)筛查检测平台,了解广东地区支气管肺炎患儿HBoV感染情况.方法 采用PCR技术筛查HBoV核蛋白(NP)基因片段,阳性标本作HBoV衣壳蛋白(VP)基因片段鉴定,并进行核酸序列和进化树分析.结果 从50例住院支气管肺炎患儿鼻咽分泌物中检测出1例HBoV,为广东地区首例,命名为GD-1株.HBoV VP基因部分序列分析与基因库中HBov VP基因序列同源性比对,除与韩国KNIH-2K6GJ2713株同源性为36%、与美国NH4549株同源性为77%外,与其他所有VP基因序列同源性均>95%,与法国株、北京株、加拿大株的同源性>98%.GD-1株HBoV部分VP基因片段与北京CZ株、加拿大株、西班牙株、意大利株等在同一基因进化簇主分枝中,与法国175/03/2002FRA株同在一个侧枝中.结论 广东地区存在HBoV感染,有进一步研究的必要性.     

山东省肠道病毒71型分离株SDLY11全基因组序列分析

目的了解2009年山东省临沂市分离的肠道病毒71型(EV71)分离株SDLY11的基因组序列特征,探讨病毒的神经毒力候选位点。[HTH]方法自手足口病患者的粪便标本中分离EV71,采用一步法RT PCR对病毒株基因组进行全序列扩增,运用DNAstar 和MEGA 4软件进行序列分析。[HTH]结果SDLY 11基因组全长为7 405 nt,其中5′非编码区（UTR） 742 nt,3′UTR 84nt,中间为6 579 nt的开放阅读框架（ORF）,编码2 193个氨基酸的多聚蛋白。VP1区及全基因组序列系统进化分析表明SDLY 11位于EV71病毒C4亚型的分支,同源性分析显示SDLY 11与安徽阜阳株同源性最高。序列比对结果发现,CNS累及HFMD患者病毒株在5[WTBX]′[WTBZ]UTR区出现了两处核苷酸位点的突变（T40C、C575T）,在VP2区第144位出现了氨基酸的突变（T144S）。[HTH]结论SDLY 11分离株属EV71病毒C4亚型。5′UTR区的两处突变（T40C、C575T）及VP2区的一处突变（T144S）可能与病毒的致神经毒力作用有关。     

2010年福建省手足口病患者中柯萨奇B5病毒的序列分析

摘要：目的 了解福建省手足口病患者中CVB5（Coxsackie virus B5）病毒的变异及进化特征。方法 采集2010年福建省手足口病例呼吸道标本,经RD细胞分离肠道病毒。病毒分离上清用于RNA提取并经RT-PCR法鉴定病毒型别。对鉴定为其他肠道病毒（非EV71或CVA16）的病毒,扩增病毒完整VP1区,扩增产物经克隆、筛选后测序。应用Mega软件对病毒VP1序列进变异比较和种系进化分析。结果 从2010年福建省手足口病患者中扩增得到6份完整的VP1区序列,序列比对证实有4份CVB5病毒。序列差异比较表明,分离自福建省的CVB5病毒一致性程度较高,在种系进化上处于独立的进化分支,而与国内其他省份或其他国家的CVB5病毒分离株比较则有较大差异。结论 2010年福建省手足口病患者中存在CVB5病毒的感染,病毒与既往其它省份分离株有较大差异,提示CVB5病毒在福建省具有独特的传播链。     

Human bocavirus infection in young children in the United States: molecular epidemiological profile and clinical characteristics of a newly emerging respiratory virus

BACKGROUND: Human bocavirus (HBoV) is a newly identified human parvovirus that was originally identified in the respiratory secretions of children with respiratory tract disease. To further investigate the epidemiological profile and clinical characteristics of HBoV infection, we screened infants and children <2 years of age (hereafter referred to as "children") for HBoV. METHODS: Children for whom respiratory specimens submitted to a diagnostic laboratory tested negative for respiratory syncytial virus, parainfluenza viruses (types 1-3), influenza A and B viruses, and adenovirus, as well as asymptomatic children, underwent screening for HBoV by use of polymerase chain reaction (PCR). Respiratory specimens were obtained from the children from 1 January 2004 through 31 December 2004. RESULTS: Twenty-two (5.2%) of the 425 children who had a respiratory specimen submitted to the diagnostic laboratory and 0 of the 96 asymptomatic children were found to be positive for HBoV by PCR (P=.02). Fever, rhinorrhea, cough, and wheezing were observed in > or =50% of the HBoV-positive children. Of the 17 children who had chest radiography performed, 12 (70.6%) had abnormal findings. HBoV appeared to have a seasonal distribution. Nucleotide polymorphisms were detected in the viral capsid protein (VP) 1/VP2 genes. Two distinct HBoV genotypes circulated during the study period. CONCLUSIONS: HBoV is circulating in the United States and is associated with both upper and lower respiratory tract disease in infants and young children.     

Clinical and molecular epidemiology of human bocavirus in respiratory and fecal samples from children in Hong Kong

BACKGROUND: Human bocavirus (HBoV) is a recently discovered parvovirus associated with respiratory tract infections in children. We conducted the first systematic prospective clinical and molecular study using nasopharyngeal aspirates (NPAs) and fecal samples. METHODS: NPAs negative for influenza virus, parainfluenza virus, respiratory syncytial virus, adenovirus, and coronavirus and fecal samples from patients with acute gastroenteritis were included. On the basis of results from a pilot study using 400 NPAs from all age groups, a prospective 12-month study was conducted to detect HBoV in 1,200 NPAs and 1,435 fecal samples from patients <18 years old by polymerase chain reaction. The complete genome sequences of HBoVs from 12 NPAs and 12 fecal samples were determined. RESULTS: Of the 400 NPAs collected in the pilot study, 20 (5.0%) were found to contain HBoV, all from children <5 years old. In the subsequent prospective study of pediatric patients, HBoV was detected in 83 (6.9%) of 1,200 NPAs. Upper and lower respiratory tract infections were equally common. HBoV was detected in 30 (2.1%) of 1,435 fecal samples. Fever and watery diarrhea were the most common symptoms. The seasonality of HBoV in NPAs and fecal samples was similar. Codetection with other pathogens occurred in 33% and 56% of NPAs and fecal samples, respectively, from patients with HBoV infection. Genomes of HBoVs from NPAs and fecal samples displayed minimal sequence variations. CONCLUSIONS: HBoV was detected in fecal specimens in children with acute gastroenteritis. A single lineage of HBoV was associated with both respiratory tract and enteric infections.     

Prevalence and Molecular Characterization of Human Bocavirus Detected in Croatian Children with Respiratory Infection

Human bocavirus (HBoV) 1 is considered an important respiratory pathogen, while the role of HBoV2-4 in clinical disease remains somewhat controversial. Since, they are characterized by a rapid evolution, worldwide surveillance of HBoVs’ genetics is necessary. This study explored the prevalence of HBoV genotypes in pediatric patients with respiratory tract infection in Croatia and studied their phylogeny. Using multiplex PCR for 15 respiratory viruses, we investigated 957 respiratory samples of children up to 18 years of age with respiratory tract infection obtained from May 2017 to March 2021 at two different hospitals in Croatia. Amplification of HBoV near-complete genome or three overlapping fragments was performed, sequenced, and their phylogenetic inferences constructed. HBoV was detected in 7.6% children with a median age of 1.36 years. Co-infection was observed in 82.2% samples. Sequencing was successfully performed on 29 HBoV positive samples, and all belonged to HBoV1. Croatian HBoV1 sequences are closely related to strains isolated worldwide, and no phylogenetic grouping based on mono- or co-infection cases or year of isolation was observed. Calculated rates of evolution for HBoV1 were 10⁻⁴ and 10⁻⁵ substitutions per site and year. Recombination was not detected among sequences from this study.     

Epidemiological profile and clinical associations of human bocavirus and other human parvoviruses

BACKGROUND: Human bocavirus (HBoV) and PARV4 are newly discovered human parvoviruses. HBoV, which was first detected in respiratory samples, has a potential role in the development of human respiratory disease. The present study compared the frequencies, epidemiological profiles, and clinical backgrounds of HBoV and PARV4 infections with those of other respiratory virus infections, by evaluating diagnostic samples referred to the Specialist Virology Laboratory (SVL) at the Royal Infirmary of Edinburgh (Edinburgh, United Kingdom). METHODS: Anonymized samples and study subject information were obtained from the respiratory sample archive of the SVL. Samples were screened for HBoV, PARV4, B19, respiratory syncytial virus (RSV), adenoviruses, influenza viruses, and parainfluenza viruses by use of nested polymerase chain reaction. RESULTS: HBoV infection was detected in 47 (8.2%) of 574 study subjects, ranking third in prevalence behind RSV infection (15.7%) and adenovirus infection (10.3%). Peak incidences of HBoV were noted among infants and young children (age, 6-24 months) during the midwinter months (December and January) and were specifically associated with lower respiratory tract infections. HBoV infections were frequently accompanied by other respiratory viruses (frequency, 43%), and they were more prevalent among individuals infected with other respiratory viruses (17%), frequently adenovirus or RSV. All respiratory samples were negative for PARV4. CONCLUSIONS: In the present study, HBoV was a frequently detected, potential respiratory pathogen, with a prevalence and an epidemiological profile comparable to those of RSV. Identification of HBoV infections may be clinically important in the future.     

Human bocavirus and human metapneumovirus in hospitalized children with lower respiratory tract illness in Changsha,China

Background

Lower respiratory tract illness is a major cause of morbidity and mortality in children worldwide, however, information about the epidemiological and clinical characteristics of LRTIs caused by HMPV and HBoV in China is limited.

Objectives

Human bocavirus (HBoV) and human metapneumovirus (HMPV) are two important viruses for children with lower respiratory tract infections (LRTI). We aimed to assay the correlation between viral load and clinical characteristics of HBoV and HMPV with LRTI in Changsha, China.

Methods

Nasopharyngeal aspirates (NPAs) from children with LRTI were collected. Real‐time PCR was used to screen HBoV and HMPV. Analyses were performed using SPSS 16.0 software.

Results

Pneumonia was the most frequent diagnosis. There was no significant difference between HBoV‐ and HMPV‐positive patients in age (P = .506) or hospitalization duration (P = .280); 24.1% and 18.2% were positive for HBoV and HMPV. HBoV infections peaked in summer (32.2%), and HMPV infections peaked in winter (28.9%). The HBoV‐positive patients had a shorter hospitalization duration than the HBoV‐negative patients (P = .021), and the HMPV‐positive patients had a higher prevalence of fever than the HMPV‐negative patients (P = .002). The HBoV viral load was significantly higher among patients aged <1 year (P = .006). The mean HBoV and HMPV viral loads were not significantly different between patients with single infections and coinfections. Patients infected with HBoV only were older than those coinfected with HBoV and other respiratory viruses (P = .005). No significant difference was found in the clinical characteristics of patients infected with HMPV only and those coinfected with HMPV and other respiratory viruses.

Conclusion

Pneumonia was the most frequent diagnosis caused by HBoV and HMPV. Neither HBoV nor HMPV viral load was correlated with disease severity.     

Identification of Human Bocavirus type 4 in a child asymptomatic for respiratory tract infection and acute gastroenteritis – Goiânia,Goiás,Brazil

Human Bocavirus (HBoV) has been identified from feces and respiratory samples from cases of both acute gastroenteritis and respiratory illness as well as in asymptomatic individuals.The aim of this study was to detect and characterize HBoV from fecal samples collected from hospitalized children aged less than five years old with no symptoms of respiratory tract infection (RTI) or acute gastroenteritis (AGE). The study involved 119 children and one fecal sample was collected from each participant between 2014 and 2015. HBoV was detected using Nested-PCR, and the viral type identified by genomic sequencing. HBoV-4 was identified from one sample obtained from a hospitalized child with soft tissue tumor of the submandibular region. This is the first report of HBoV-4 identification in Brazil, but we consider that this type may be circulating in the country similar to the other types and new investigations are necessary.     

High prevalence of human bocavirus 1 in infants with lower acute respiratory tract disease in Argentina, 2007-2009

Human bocavirus (HBoV) is a parvovirus whose association with respiratory disease is currently under investigation.ObjectiveTo determine HBoV prevalence in children with lower acute respiratory infection.MethodsWe investigated HBoV in 433 nasopharyngeal aspirates collected in 2007–2009 from children 0 to 5 years old hospitalized with bronchiolitis or pneumonia in Córdoba, Argentina.ResultsThe general prevalence of HBoV was 21.5% and the positive cases (HBoV+) were more frequent during winter and spring. The mean age of HBoV+ patients was 6.9 months, with 87.1% of the detections corresponding to infants less than 1 year old (among which the prevalence of HBoV was 26.3% in patients < 3 months of age, 22.1% in 3 to 6 months, 25.3% in 6 to 9 months, and 18.8% in 9 to 12 months). The sequence analysis of the NP1 coding region of 15 isolates showed that all isolates from Cordoba were HBoV1 which exhibited a homology of nearly 100% both among themselves and with the originally discovered virus from 2005.ConclusionOverall, our results indicate that HBoV is a significant pathogen that contributes to acute respiratory infection both on its own and during coinfection with other viruses.     

Human bocavirus amongst an all‐ages population hospitalised with acute lower respiratory infections in Cambodia

Please cite this paper as: Arnott et al. (2013) Human bocavirus amongst an all‐ages population hospitalised with acute lower respiratory infections in Cambodia. Influenza and Other Respiratory Viruses 7(2) 201–210. Background Human bocavirus (HBoV) is a novel parvovirus that is associated with respiratory and gastrointestinal tract disease. Objectives To investigate the prevalence and genetic diversity of HBoV amongst hospitalized patients with acute lower respiratory infection (ALRI) in Cambodia. Study Design Samples were collected from 2773 patients of all ages hospitalised with symptoms of ALRI between 2007 and 2009. All samples were screened by multiplex RT‐PCR/PCR for 18 respiratory viruses. All samples positive for HBoV were sequenced and included in this study. Results Of the samples tested, 43 (1·5%) were positive for HBoV. The incidence of HBoV did not vary between the consecutive seasons investigated, and HBoV infections were detected year‐round. The incidence of HBoV infection was highest in patients aged <2 years, with pneumonia or bronchopneumonia the most common clinical diagnosis, regardless of age. A total of 19 patients (44%) were co‐infected with HBoV and an additional respiratory pathogen. All isolates were classified as HBoV type 1 (HBoV‐1). High conservation between Cambodian NP1 and V1V2 gene sequences was observed. Conclusions Human bocavirus infection can result in serious illness, however is frequently detected in the context of viral co‐infection. Specific studies are required to further understand the true pathogenesis of HBoV in the context of severe respiratory illness.