Real-Time Quantitative PCR Detection of Four Human Bocaviruses
- 1 November 2010
- journal article
- research article
- Published by American Society for Microbiology in Journal of Clinical Microbiology
- Vol. 48 (11), 4044-4050
- https://doi.org/10.1128/jcm.00686-10
Abstract
Human bocavirus (HBoV) was discovered in 2005 and is associated with respiratory tract symptoms in young children. Three additional members of the genus B ocavirus , HBoV2, -3, and -4, were discovered recently from fecal specimens, and early results indicate an association between HBoV2 and gastrointestinal disease. In this study, we present an undifferentiating multiplex real-time quantitative PCR assay for the detection of these novel viruses. Differentiation of the individual bocavirus species can be subsequently achieved with corresponding singleplex PCRs or by sequencing. Both multiplex and singleplex assays were consistently able to detect ≤10 copies of HBoV1 to -4 plasmid templates/reaction, with dynamic quantification ranges of 8 logs and 97% to 102% average reaction efficiencies. These new assays were used to screen stool samples from 250 Finnish patients (median age, 40 years) that had been sent for diagnosis of gastrointestinal infection. Four patients (1.6%; median age, 1.1 years) were reproducibly positive for HBoV2, and one patient (0.4%; 18 years of age) was reproducibly positive for HBoV3. The viral DNA loads varied from 3 to 10 9 copies/ml of stool extract. None of the stool samples harbored HBoV1 or HBoV4. The highly conserved sequence of the hydrolysis probe used in this assay may provide a flexible future platform for the quantification of additional, hitherto-unknown human bocaviruses that might later be discovered. Our results support earlier findings that HBoV2 is a relatively common pathogen in the stools of diarrheic young children, yet does not often occur in the stools of adults.Keywords
This publication has 22 references indexed in Scilit:
- Human bocavirus species 2 and 3 in BrazilJournal of Clinical Virology, 2010
- Serodiagnosis of Human Bocavirus InfectionClinical Infectious Diseases, 2008
- Real-Time PCR for Diagnosis of Human Bocavirus Infections and Phylogenetic AnalysisJournal of Clinical Microbiology, 2007
- Human Bocavirus and Acute Wheezing in ChildrenClinical Infectious Diseases, 2007
- Human Bocavirus: A Novel Parvovirus Epidemiologically Associated with Pneumonia Requiring Hospitalization in ThailandThe Journal of Infectious Diseases, 2007
- Human bocavirus in Italian patients with respiratory diseasesJournal of Clinical Virology, 2007
- Human Bocavirus Infection in Young Children in the United States: Molecular Epidemiological Profile and Clinical Characteristics of a Newly Emerging Respiratory VirusThe Journal of Infectious Diseases, 2006
- Real-Time PCR Assays for Detection of Bocavirus in Human SpecimensJournal of Clinical Microbiology, 2006
- Cloning of a human parvovirus by molecular screening of respiratory tract samplesProceedings of the National Academy of Sciences of the United States of America, 2005
- Mfold web server for nucleic acid folding and hybridization predictionNucleic Acids Research, 2003