{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T19:15:11Z","timestamp":1791314111731,"version":"4.3.1"},"reference-count":45,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2021,8,31]],"date-time":"2021-08-31T00:00:00Z","timestamp":1630368000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004488","name":"Hrvatska Zaklada za Znanost","doi-asserted-by":"publisher","award":["IP-2016-06-7556"],"award-info":[{"award-number":["IP-2016-06-7556"]}],"id":[{"id":"10.13039\/501100004488","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>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\u2019 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\u22124 and 10\u22125 substitutions per site and year. Recombination was not detected among sequences from this study.<\/jats:p>","DOI":"10.3390\/v13091728","type":"journal-article","created":{"date-parts":[[2021,9,1]],"date-time":"2021-09-01T03:13:17Z","timestamp":1630465997000},"page":"1728","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["Prevalence and Molecular Characterization of Human Bocavirus Detected in Croatian Children with Respiratory Infection"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6405-6922","authenticated-orcid":false,"given":"Sun\u010danica","family":"Ljubin-Sternak","sequence":"first","affiliation":[{"name":"Molecular Microbiology Department, Dr. Andrija \u0160tampar Teaching Institute of Public Health, 10000 Zagreb, Croatia"},{"name":"Medical Microbiology Department, University of Zagreb School of Medicine, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anamarija","family":"Slovi\u0107","sequence":"additional","affiliation":[{"name":"Center of Excellence for Virus Immunology and Vaccines, Center for Research and Knowledge Transfer in Biotechnology, University of Zagreb, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maja","family":"Mija\u010d","sequence":"additional","affiliation":[{"name":"Molecular Microbiology Department, Dr. Andrija \u0160tampar Teaching Institute of Public Health, 10000 Zagreb, Croatia"},{"name":"Medical Microbiology Department, University of Zagreb School of Medicine, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mirna","family":"Jurkovi\u0107","sequence":"additional","affiliation":[{"name":"Center of Excellence for Virus Immunology and Vaccines, Center for Research and Knowledge Transfer in Biotechnology, University of Zagreb, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dubravko","family":"For\u010di\u0107","sequence":"additional","affiliation":[{"name":"Center of Excellence for Virus Immunology and Vaccines, Center for Research and Knowledge Transfer in Biotechnology, University of Zagreb, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irena","family":"Ivkovi\u0107-Jurekovi\u0107","sequence":"additional","affiliation":[{"name":"Department of Pulmonology, Allergy, Immunology and Rheumatology, Children\u2019s Hospital Zagreb, 10000 Zagreb, Croatia"},{"name":"Faculty for Dental Medicine and Healthcare\/School of Medicine, Josip Juraj Strossmayer University of Osijek, 31000 Osijek, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4056-7880","authenticated-orcid":false,"given":"Tatjana","family":"Tot","sequence":"additional","affiliation":[{"name":"Microbiology Department, General hospital Karlovac, Karlovac, 47000 Karlovac, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jasmina","family":"Vrane\u0161","sequence":"additional","affiliation":[{"name":"Molecular Microbiology Department, Dr. Andrija \u0160tampar Teaching Institute of Public Health, 10000 Zagreb, Croatia"},{"name":"Medical Microbiology Department, University of Zagreb School of Medicine, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,8,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"12891","DOI":"10.1073\/pnas.0504666102","article-title":"Cloning of a human parvovirus by molecular screening of respiratory tract samples","volume":"102","author":"Allander","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"264","DOI":"10.3201\/eid1802.111293","article-title":"Association of human bocavirus 1 infection with respiratory disease in childhood follow-up study, Finland","volume":"18","author":"Meriluoto","year":"2012","journal-title":"Emerg. Infect. Dis."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1016\/j.jcv.2010.07.016","article-title":"Human bocavirus in children: Mono-detection, high viral load and viraemia are associated with respiratory tract infection","volume":"49","author":"Christensen","year":"2010","journal-title":"J. Clin. Virol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Arthur, J.L., Higgins, G.D., Davidson, G.P., Givney, R.C., and Ratcliff, R.M. (2009). A novel bocavirus associated with acute gastroenteritis in Australian children. PLoS Pathog., 5.","DOI":"10.1371\/journal.ppat.1000391"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1633","DOI":"10.1086\/652416","article-title":"Human bocaviruses are highly diverse, dispersed, recombination prone, and prevalent in enteric infections","volume":"201","author":"Kapoor","year":"2010","journal-title":"J. Infect. Dis."},{"key":"ref_6","unstructured":"International Committee on Taxonomy of Viruses (ICTV) (2021, July 04). Virus Taxonomy: 2017. Available online: https:\/\/talk.ictvonline.org\/taxonomy."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"L\u00fcsebrink, J., Schildgen, V., Tillmann, R.L., Wittleben, F., B\u00f6hmer, A., M\u00fcller, A., and Schildgen, O. (2011). Detection of Head-to-Tail DNA Sequences of Human Bocavirus in Clinical Samples. PloS ONE, 6.","DOI":"10.1371\/journal.pone.0019457"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Huang, Q., Deng, X., Yan, Z., Cheng, F., Luo, Y., Shen, W., Lei-Butters, D.C., Chen, A.Y., Li, Y., and Tang, L. (2012). Establishment of a reverse genetics system for studying human bocavirus in human airway epithelia. PLoS Pathog., 8.","DOI":"10.1371\/journal.ppat.1002899"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"696604","DOI":"10.3389\/fmicb.2021.696604","article-title":"Recent Advances in Molecular Biology of Human Bocavirus 1 and Its Applications","volume":"12","author":"Shao","year":"2021","journal-title":"Front. Microbiol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1016\/j.meegid.2009.11.011","article-title":"Rapid molecular evolution of human bocavirus revealed by Bayesian coalescent inference","volume":"10","author":"Zehender","year":"2010","journal-title":"Infect. Genet. Evol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1016\/j.meegid.2013.03.040","article-title":"Complete genome sequence analysis of novel human bocavirus reveals genetic recombination between human bocavirus 2 and human bocavirus 4","volume":"17","author":"Khamrin","year":"2013","journal-title":"Infect. Genet. Evol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"382","DOI":"10.1016\/j.meegid.2014.08.022","article-title":"Prevalence analysis of different human bocavirus genotypes in pediatric patients revealed intra-genotype recombination","volume":"27","author":"Zhao","year":"2014","journal-title":"Infect. Genet. Evol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2737","DOI":"10.3389\/fmicb.2019.02737","article-title":"The Emerging Role of Rhinoviruses in Lower Respiratory Tract Infections in Children-Clinical and Molecular Epidemiological Study from Croatia, 2017\u20132019","volume":"10","author":"Tot","year":"2019","journal-title":"Front. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1111\/j.1750-2659.2012.00360.x","article-title":"The impact of bacterial and viral co-infection in severe influenza","volume":"7","author":"Blyth","year":"2013","journal-title":"Influenza Other Respir. Viruses"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1038\/nmeth.1923","article-title":"Fast gapped-read alignment with Bowtie 2","volume":"9","author":"Langmead","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2078","DOI":"10.1093\/bioinformatics\/btp352","article-title":"The Sequence Alignment\/Map format and SAMtools","volume":"25","author":"Li","year":"2009","journal-title":"Bioinformatics"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"821","DOI":"10.1101\/gr.074492.107","article-title":"Velvet: Algorithms for de novo short read assembly using de Bruijn graphs","volume":"18","author":"Zerbino","year":"2008","journal-title":"Genome. Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"772","DOI":"10.1093\/molbev\/mst010","article-title":"MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability","volume":"30","author":"Katoh","year":"2013","journal-title":"Mol. Biol. Evol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"3276","DOI":"10.1093\/bioinformatics\/btu531","article-title":"AliView: A fast and lightweight alignment viewer and editor for large datasets","volume":"30","author":"Larsson","year":"2014","journal-title":"Bioinformatics"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2725","DOI":"10.1093\/molbev\/mst197","article-title":"MEGA6: Molecular evolutionary genetics analysis version 6.0","volume":"30","author":"Tamura","year":"2013","journal-title":"Mol. Biol. Evol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"722","DOI":"10.1038\/nmeth.2109","article-title":"JModelTest 2: More models, new heuristics and parallel computing","volume":"9","author":"Darriba","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1969","DOI":"10.1093\/molbev\/mss075","article-title":"Bayesian phylogenetics with BEAUti and the BEAST 1.7","volume":"29","author":"Drummond","year":"2012","journal-title":"Mol. Biol. Evol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"vev003","DOI":"10.1093\/ve\/vev003","article-title":"RDP4: Detection and analysis of recombination patterns in virus genomes","volume":"1","author":"Martin","year":"2015","journal-title":"Virus Evol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4658","DOI":"10.1128\/JVI.02964-15","article-title":"Nonstructural Protein NP1 of Human Bocavirus 1 Plays a Critical Role in the Expression of Viral Capsid Proteins","volume":"90","author":"Zou","year":"2016","journal-title":"J. Virol."},{"key":"ref_25","first-page":"65","article-title":"High Detection Rates of Human Bocavirus in Infants and Small Children with Lower Respiratory Tract Infection from Croatia","volume":"1","author":"Tabain","year":"2019","journal-title":"Clin. Lab."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Polo, D., Lema, A., G\u00e1ndara, E., and Romalde, J.L. (2021). Prevalence of human bocavirus infections in Europe. A systematic review and meta-analysis. Transbound. Emerg. Dis., Epub ahead of print.","DOI":"10.1111\/tbed.14233"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"8684","DOI":"10.3748\/wjg.v22.i39.8684","article-title":"Human bocavirus: Current knowledge and future challenges","volume":"22","author":"Guido","year":"2016","journal-title":"World J. Gastroenterol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1625","DOI":"10.1086\/652405","article-title":"Frequent and Prolonged Shedding of Bocavirus in Young Children Attending Daycare","volume":"201","author":"Martin","year":"2010","journal-title":"J. Infect. Dis."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1423","DOI":"10.3201\/eid1509.090204","article-title":"Clinical assessment and improved diagnosis of bocavirus-induced wheezing in children, Finland","volume":"15","author":"Lahtinen","year":"2009","journal-title":"Emerg. Infect. Dis."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"418","DOI":"10.1016\/S2352-4642(19)30057-4","article-title":"Human bocaviruses and paediatric infections","volume":"3","author":"Christensen","year":"2019","journal-title":"Lancet Child Adolesc. Health"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1186\/s12879-018-3225-3","article-title":"Epidemiology of HBoV1 infection and relationship with meteorological conditions in hospitalized pediatric patients with acute respiratory illness: A 7-year study in a subtropical region","volume":"18","author":"Liu","year":"2018","journal-title":"BMC Infect. Dis."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2011","DOI":"10.1002\/jmv.23880","article-title":"Human bocavirus isolated from children with acute respiratory tract infections in Korea, 2010\u20132011","volume":"86","author":"Ahn","year":"2014","journal-title":"J. Med. Virol."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Kenmoe, S., Vernet, M.A., Njankouo-Ripa, M., Penlap, V.B., Vabret, A., and Njouom, R. (2017). Phylogenic analysis of human bocavirus detected in children with acute respiratory infection in Yaounde, Cameroon. BMC Res. Notes, 10.","DOI":"10.1186\/s13104-017-2620-y"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1005","DOI":"10.1099\/jmm.0.001219","article-title":"Human bocavirus (HBoV) in Kuwait: Molecular epidemiology and clinical outcome of the virus among patients with respiratory diseases","volume":"69","author":"Madi","year":"2020","journal-title":"J. Med. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2118","DOI":"10.1128\/JCM.00098-12","article-title":"Detection of human bocaviruses 1 to 4 from nasopharyngeal swab samples collected from patients with respiratory tract infections","volume":"50","author":"Koseki","year":"2012","journal-title":"J. Clin. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1698","DOI":"10.3201\/eid1510.090337","article-title":"Human bocavirus 2 in children, South Korea","volume":"15","author":"Han","year":"2009","journal-title":"Emerg. Infect. Dis."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1007\/s10096-014-2215-7","article-title":"Epidemic and molecular evolution of human bocavirus in hospitalized children with acute respiratory tract infection","volume":"34","author":"Lu","year":"2015","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/j.jinf.2016.05.014","article-title":"Genetic characterization of human bocavirus among children with severe acute respiratory infection in China","volume":"73","author":"Wang","year":"2016","journal-title":"J. Infect."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1111\/imm.12202","article-title":"Immunogenicity of recombinant human bocavirus-1,2 VP2 gene virus-like particles in mice","volume":"142","author":"Deng","year":"2014","journal-title":"Immunology"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1016\/S0969-2126(03)00152-7","article-title":"Crystal structure of the SF3 helicase from adeno-associated virus type 2","volume":"11","author":"James","year":"2003","journal-title":"Structure"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"7974","DOI":"10.1128\/jvi.68.12.7974-7985.1994","article-title":"Targeting of promoters for trans activation by a carboxy-terminal domain of the NS-1 protein of the parvovirus minute virus of mice","volume":"68","author":"Legendre","year":"1994","journal-title":"J. Virol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1016\/j.bbrc.2007.10.164","article-title":"Phospholipase A2-like activity of human bocavirus VP1 unique region","volume":"365","author":"Qu","year":"2008","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1016\/S1534-5807(01)00031-4","article-title":"A viral phospholipase A2 is required for parvovirus infectivity","volume":"1","author":"Zadori","year":"2001","journal-title":"Dev. Cell."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"4670","DOI":"10.1128\/JVI.02998-15","article-title":"Mapping antigenic epitopes on the human bocavirus capsid","volume":"90","author":"Kailasan","year":"2016","journal-title":"J. Virol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.meegid.2014.08.026","article-title":"Recombination in the evolution of human bocavirus","volume":"28","author":"Tyumentsev","year":"2014","journal-title":"Infect. Genet. Evol."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/9\/1728\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:53:00Z","timestamp":1760165580000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/9\/1728"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,8,31]]},"references-count":45,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2021,9]]}},"alternative-id":["v13091728"],"URL":"https:\/\/doi.org\/10.3390\/v13091728","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,8,31]]}}}