Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2011 Apr;49(4):1537-41.
doi: 10.1128/JCM.00196-10. Epub 2011 Feb 16.

Development of a real-time PCR assay for detecting and quantifying human bocavirus 2

Zi-qian Xu  1 , Wei-xia Cheng, Bo-wen Li, Jie Li, Bei Lan, Zhao-jun Duan
Affiliations

Development of a real-time PCR assay for detecting and quantifying human bocavirus 2

Zi-qian Xu et al. J Clin Microbiol. 2011 Apr.

Abstract

Human bocavirus 2 (HBoV2) is a parvovirus that has been recently identified in stool samples from children. Any association between the virus and clinical disease is unclear. A rapid, reliable diagnostic method is necessary to address this issue. In this study, we developed a sensitive and specific HBoV2 quantitative real-time PCR assay that targets the HBoV2 NP-1 gene, based on the TaqMan method. The assay could reproducibly detect 10 copies of a recombinant DNA plasmid containing a partial region of the HBoV2 genome, with a dynamic range of 8 log units (10(1) to 10(8) copies). A clinical evaluation detected HBoV2 in 85 (24.6%) of 345 children with gastroenteritis, with viral loads ranging from 1.67 × 10(2) to 4.27 × 10(9) copies per ml of stool specimen.

PubMed Disclaimer

Figures

Fig. 1.
Fig. 1.
NP-1 real-time PCR assay showing representative results obtained with serial 10-fold dilutions (101 to 108 copies per reaction) of the pGEM-T NP-1 HBoV2 plasmid, run in triplicate. The left panel shows baseline subtractive curve fit views of the data with relative fluorescence units (RFU) plotted against cycle number. The right panel shows a standard curve analysis of the DNA amplification plots with cycle threshold (CT) values plotted against the logarithm of the input copy number. The dynamic range of the NP-1 PCR assay spanned 8 log units with a slope of −3.3 and an R2 value of 1.0.
Fig. 2.
Fig. 2.
Viral loads of HBoV2 in stools according to the real-time PCR assay. The median viral load of 38 positive specimens with both assays was 3.37 × 104 copies/ml, ranging from 1.93 × 103 to 4.27 × 109copies per ml. The median viral load of 47 positive specimens only by real-time PCR was 4.01 × 103 copies/ml, ranging from 1.67 × 102 to 8.76 × 104 copies per ml.
Fig. 3.
Fig. 3.
Seasonal distribution of the children hospitalized with acute gastroenteritis, November 2007 to October 2008, at Taiyuan Children Hospital. The numbers of cases (bars, left y axis) and the HBoV2-positive rate (line, right y axis) varied in different months (x axis). “total” means the number of all of the cases. “HBoV2” means the number of HBoV2-positive cases.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Allander T., et al. 2007. Human bocavirus and acute wheezing in children. Clin. Infect. Dis. 44:904–910 - PMC - PubMed
    1. Allander T., et al. 2005. Cloning of a human parvovirus by molecular screening of respiratory tract samples. Proc. Natl. Acad. Sci. U. S. A. 102:12891–12896 - PMC - PubMed
    1. Arthur J. L., Higgins G. D., Davidson G. P., Givney R. C., Ratcliff R. M. 2009. A novel bocavirus associated with acute gastroenteritis in Australian children. PLoS Pathog. 5:e1000391. - PMC - PubMed
    1. Cecchini S., et al. 2009. Evidence of prior exposure to human bocavirus: a retrospective serological study of 404 adult sera in the United States. Clin. Vaccine Immunol. 16:597–604 - PMC - PubMed
    1. Chan M. C. W., et al. 2006. Fecal viral load and Norovirus-associated gastroenteritis. Emerg. Infect. Dis. 12:1278–1280 - PMC - PubMed

Publication types

Associated data