Abstract
Human bocavirus (HBoV) has been detected worldwide in respiratory samples. Two real‐time PCR assays, targeting the non‐structural protein (NP‐1) and viral protein (VP‐1) genes, were designed and validated to detect HBoV in patients with respiratory disease, gastroenteritis, or systemic illness. Sensitivity of the NP‐1 and VP‐1 assays were equal to the conventional PCR assay previously described by Allander et al. [2005: Proc Natl Acad Sci USA 102: 12891–12896] being 100%, and giving specificity of 94% and 93%, respectively. There was no cross‐reaction identified with unrelated respiratory agents, or to human DNA. The limits of detection were 10 copies of genomic DNA equivalents per reaction for both assays. The assays were used to screen three different sample populations, combined nose, and throat swabs (n = 96) from children with acute respiratory disease, fecal samples (n = 375) from adults, and children with gastroenteritis and whole blood (n = 229) collected from 31 immunocompromised children taken over an 18‐month period. In total 17 (18%) respiratory samples and 18 (4.8%) fecal samples were identified as having HBoV present. Of the pediatric whole blood specimens investigated, HBoV was detected in six (2.6%) samples from four patients. In summary, two real‐time PCR assays targeting different genes were designed and validated for use as screening methods for the detection of HBoV. HBoV was found in three different specimen types: parent‐collected combined nose–throat swabs, fecal samples collected from symptomatic individuals and whole blood from immunocompromised children. J. Med. Virol. 81:488–493, 2009. © 2009 Wiley‐Liss, Inc.
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This study was conducted exclusively at the Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital and Health Service, Brisbane, Queensland, Australia.
REFERENCES
- Allander T, Tammi MT, Eriksson M, Bjerkner A, Tiveljung‐Lindell A, Andersson B. 2005. Cloning of a human parvovirus by molecular screening of respiratory tract samples. Proc Natl Acad Sci USA 102: 12891–12896. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Allander T, Jartti T, Gupta S, Niesters HG, Lehtinen P, Osterback R, Vuorinen T, Waris M, Bjerkner A, Tiveljung‐Lindell A, van den Hoogen BG, Hyypia T, Ruuskanen O. 2007. Human bocavirus and acute wheezing in children. Clin Infect Dis 44: 904–910. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Allard A, Albinsson B, Wadell G. 1992. Detection of adenoviruses in stools from healthy persons and patients with diarrhea by two‐step polymerase chain reaction. J Med Virol 37: 149–157. [DOI] [PubMed] [Google Scholar]
- Arden KE, McErlean P, Nissen MD, Sloots TP, Mackay IM. 2006. Frequent detection of human rhinoviruses, paramyxoviruses, coronaviruses, and bocavirus during acute respiratory tract infections. J Med Virol 78: 1232–1240. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Arnold JC, Singh KK, Spector SA, Sawyer MH. 2006. Human bocavirus: Prevalence and clinical spectrum at a children's hospital. Clin Infect Dis 43: 283–288. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bastien N, Brandt K, Dust K, Ward D, Li Y. 2006. Human bocavirus infection, Canada. Emerg Infect Dis 12: 848–850. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bialasiewicz S, Whiley DM, Lambert SB, Gould A, Nissen MD, Sloots TP. 2007. Development and evaluation of real‐time PCR assays for the detection of the newly identified KI and WU polyomaviruses. J Clin Virol 40: 9–14. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carmichael LE, Joubert JC, Pollock RV. 1980. Hemagglutination by canine parvovirus: Serologic studies and diagnostic applications. Am J Vet Res 41: 784–791. [PubMed] [Google Scholar]
- Chieochansin T, Samransamruajkit R, Chutinimitkul S, Payungporn S, Hiranras T, Theamboonlers A, Poovorawan Y. 2008. Human bocavirus (HBoV) in Thailand: Clinical manifestations in a hospitalized pediatric patient and molecular virus characterization. J Infect 56: 137–142. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Choi EH, Lee HJ, Kim SJ, Eun BW, Kim NH, Lee JA, Lee JH, Song EK, Kim SH, Park JY, Sung JY. 2006. The association of newly identified respiratory viruses with lower respiratory tract infections in Korean children, 2000–2005. Clin Infect Dis 43: 585–592. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Durham PJ, Lax A, Johnson RH. 1985. Pathological and virological studies of experimental parvoviral enteritis in calves. Res Vet Sci 38: 209–219. [PubMed] [Google Scholar]
- Esposito S, Lizioli A, Lastrico A, Begliatti E, Rognoni A, Tagliabue C, Cesati L, Carreri V, Principi N. 2007. Impact on respiratory tract infections of heptavalent pneumococcal conjugate vaccine administered at 3,5 and 11 months of age. Respir Res 8: 12. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fredericks DN, Relman DA. 1996. Sequence‐based identification of microbial pathogens: A reconsideration of Koch's postulates. Clin Microbiol Rev 9: 18–33. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fry AM, Lu X, Chittaganpitch M, Peret T, Fischer J, Dowell SF, Anderson LJ, Erdman D, Olsen SJ. 2007. Human bocavirus: A novel parvovirus epidemiologically associated with pneumonia requiring hospitalization in Thailand. J Infect Dis 195: 1038–1045. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gouvea V, Glass RI, Woods P, Taniguchi K, Clark HF, Forrester B, Fang ZY. 1990. Polymerase chain reaction amplification and typing of rotavirus nucleic acid from stool specimens. J Clin Microbiol 28: 276–282. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hall TA. 1999. Bioedit: A user‐friendly biological sequence alignment editor and analysis program for windows 95/98/NT. Nucl Acids Symp Ser 41: 95–98. [Google Scholar]
- Hirsch HH, Mohaupt M, Klimkait T. 2001. Prospective monitoring of BK virus load after discontinuing sirolimus treatment in a renal transplant patient with BK virus nephropathy. J Infect Dis 184: 1494–1495, author reply 1495–1496. [DOI] [PubMed] [Google Scholar]
- Kageyama T, Kojima S, Shinohara M, Uchida K, Fukushi S, Hoshino FB, Takeda N, Katayama K. 2003. Broadly reactive and highly sensitive assay for Norwalk‐like viruses based on real‐time quantitative reverse transcription‐PCR. J Clin Microbiol 41: 1548–1557. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kesebir D, Vazquez M, Weibel C, Shapiro ED, Ferguson D, Landry ML, Kahn JS. 2006. Human bocavirus infection in young children in the United States: Molecular epidemiological profile and clinical characteristics of a newly emerging respiratory virus. J Infect Dis 194: 1276–1282. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kupfer B, Vehreschild J, Cornely O, Kaiser R, Plum G, Viazov S, Franzen C, Tillmann RL, Simon A, Muller A, Schildgen O. 2006. Severe pneumonia and human bocavirus in adult. Emerg Infect Dis 12: 1614–1616. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lambert SB, Allen KM, Druce JD, Birch CJ, Mackay IM, Carlin JB, Carapetis JR, Sloots TP, Nissen MD, Nolan TM. 2007. Community epidemiology of human metapneumovirus, human coronavirus NL63, and other respiratory viruses in healthy preschool‐aged children using parent‐collected specimens. Pediatrics 120: e929–e937. [DOI] [PubMed] [Google Scholar]
- Lau SK, Yip CC, Que TL, Lee RA, Au‐Yeung RK, Zhou B, So LY, Lau YL, Chan KH, Woo PC, Yuen KY. 2007. Clinical and molecular epidemiology of human bocavirus in respiratory and fecal samples from children in Hong Kong. J Infect Dis 196: 986–993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lipson SM. 2002. Rapid laboratory diagnostics during the winter respiratory virus season. J Clin Microbiol 40: 733–734. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ma X, Endo R, Ishiguro N, Ebihara T, Ishiko H, Ariga T, Kikuta H. 2006. Detection of human bocavirus in Japanese children with lower respiratory tract infections. J Clin Microbiol 44: 1132–1134. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maggi F, Andreoli E, Pifferi M, Meschi S, Rocchi J, Bendinelli M. 2007. Human bocavirus in Italian patients with respiratory diseases. J Clin Virol 38: 321–325. [DOI] [PubMed] [Google Scholar]
- Manning A, Russell V, Eastick K, Leadbetter GH, Hallam N, Templeton K, Simmonds P. 2006. Epidemiological profile and clinical associations of human bocavirus and other human parvoviruses. J Infect Dis 194: 1283–1290. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Naghipour M, Cuevas LE, Bakhshinejad T, Dove W, Hart CA. 2007. Human bocavirus in Iranian children with acute respiratory infections. J Med Virol 79: 539–543. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Noel JS, Lee TW, Kurtz JB, Glass RI, Monroe SS. 1995. Typing of human astroviruses from clinical isolates by enzyme immunoassay and nucleotide sequencing. J Clin Microbiol 33: 797–801. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pal A, Sirota L, Maudru T, Peden K, Lewis AM Jr. 2006. Real‐time, quantitative PCR assays for the detection of virus‐specific DNA in samples with mixed populations of polyomaviruses. J Virol Methods 135: 32–42. [DOI] [PubMed] [Google Scholar]
- Qu XW, Duan ZJ, Qi ZY, Xie ZP, Gao HC, Liu WP, Huang CP, Peng FW, Zheng LS, Hou YD. 2007. Human bocavirus infection, People's Republic of China. Emerg Infect Dis 13: 165–168. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schenk T, Strahm B, Kontny U, Hufnagel M, Neumann‐Haefelin D, Falcone V. 2007. Disseminated bocavirus infection after stem cell transplant. Emerg Infect Dis 13: 1425–1427. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sloots TP, McErlean P, Speicher DJ, Arden KE, Nissen MD, Mackay IM. 2006. Evidence of human coronavirus HKU1 and human bocavirus in Australian children. J Clin Virol 35: 99–102. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smuts H, Hardie D. 2006. Human bocavirus in hospitalized children, South Africa. Emerg Infect Dis 12: 1457–1458. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Whiley DM, Lambert SB, Bialasiewicz S, Goire N, Nissen MD, Sloots TP. 2008. False‐negative results in nucleicacid amplification tests—Do we need to routinely use two genetic targets in all assays to overcome problems caused by sequence variation? Clit Rev Microbiol 34: 71–76. [DOI] [PubMed] [Google Scholar]