title: Real-time PCR assays for detection of bocavirus in human specimens.
abstract: The recently discovered human bocavirus (HBoV) is the first member of the family Parvoviridae, genus Bocavirus, to be potentially associated with human disease. Several studies have identified HBoV in respiratory specimens from children with acute respiratory disease, but the full spectrum of clinical disease and the epidemiology of HBoV infection remain unclear. The availability of rapid and reliable molecular diagnostics would therefore aid future studies of this novel virus. To address this, we developed two sensitive and specific real-time TaqMan PCR assays that target the HBoV NS1 and NP-1 genes. Both assays could reproducibly detect 10 copies of a recombinant DNA plasmid containing a partial region of the HBoV genome, with a dynamic range of 8 log units (10(1) to 10(8) copies). Eight blinded clinical specimen extracts positive for HBoV by an independent PCR assay were positive by both real-time assays. Among 1,178 NP swabs collected from hospitalized pneumonia patients in Sa Kaeo Province, Thailand, 53 (4.5%) were reproducibly positive for HBoV by one or both targets. Our data confirm the possible association of HBoV infection with pneumonia and demonstrate the utility of these real-time PCR assays for HBoV detection.
authors.affiliation: Respiratory and Gastroenteritis Viruses Branch, Centers for Disease Control and Prevention, 1600 Clifton Road, N.E., Mailstop G04, Atlanta, GA 30333, USA.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Lu
authors.forename: Malinee
authors.initials: M
authors.lastname: Chittaganpitch
authors.forename: Sonja J
authors.initials: SJ
authors.lastname: Olsen
authors.forename: Ian M
authors.initials: IM
authors.lastname: Mackay
authors.forename: Theo P
authors.initials: TP
authors.lastname: Sloots
authors.forename: Alicia M
authors.initials: AM
authors.lastname: Fry
authors.forename: Dean D
authors.initials: DD
authors.lastname: Erdman
mesh_terms: D004279:DNA, Viral / Q000032:analysis; D006801:Humans; D019590:Nucleocapsid Proteins / Q000235:genetics; D010322:Parvoviridae Infections / Q000821:virology*; D019219:Parvovirinae / Q000235:genetics / Q000302:isolation & purification*; D011024:Pneumonia, Viral / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019914:Taq Polymerase; D013785:Thailand; D017361:Viral Nonstructural Proteins / Q000235:genetics
chemical_list: D004279:DNA, Viral; D019590:Nucleocapsid Proteins; D017361:Viral Nonstructural Proteins; D019914:Taq Polymerase