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Journal of Clinical Microbiology, July 2007, p. 2116-2122, Vol. 45, No. 7
0095-1137/07/$08.00+0     doi:10.1128/JCM.00027-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Real-Time PCR for Diagnosis of Human Bocavirus Infections and Phylogenetic Analysis{triangledown}

Florian Neske,1 Kerstin Blessing,2 Franz Tollmann,1 Jörg Schubert,1 Axel Rethwilm,1 Hans Wolfgang Kreth,2 and Benedikt Weissbrich1*

University of Würzburg, Institute of Virology and Immunobiology, Versbacher Str. 7, 97078 Würzburg, Germany,1 University of Würzburg, Children's Hospital, Josef-Schneider-Str. 2, 97080 Würzburg, Germany2

Received 4 January 2007/ Returned for modification 19 March 2007/ Accepted 20 April 2007

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 103 copies/ml (range, 2.7 x 10° to 1.5 x 1011 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.

* Corresponding author. Mailing address: Institute of Virology and Immunobiology, University of Würzburg, Versbacher Str. 7, D-97078 Würzburg, Germany. Phone: 49 931 201 49962. Fax: 49 931 201 49561. E-mail: weissbrich{at}vim.uni-wuerzburg.de

{triangledown} Published ahead of print on 2 May 2007.

Journal of Clinical Microbiology, July 2007, p. 2116-2122, Vol. 45, No. 7
0095-1137/07/$08.00+0     doi:10.1128/JCM.00027-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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