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Optimization of HPV DNA detection in urine by improving collection, storage, and extraction

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European Journal of Clinical Microbiology & Infectious Diseases Aims and scope Submit manuscript

Abstract

The benefits of using urine for the detection of human papillomavirus (HPV) DNA have been evaluated in disease surveillance, epidemiological studies, and screening for cervical cancers in specific subgroups. HPV DNA testing in urine is being considered for important purposes, notably the monitoring of HPV vaccination in adolescent girls and young women who do not wish to have a vaginal examination. The need to optimize and standardize sampling, storage, and processing has been reported.

In this paper, we examined the impact of a DNA-conservation buffer, the extraction method, and urine sampling on the detection of HPV DNA and human DNA in urine provided by 44 women with a cytologically normal but HPV DNA-positive cervical sample. Ten women provided first-void and midstream urine samples. DNA analysis was performed using real-time PCR to allow quantification of HPV and human DNA.

The results showed that an optimized method for HPV DNA detection in urine should (a) prevent DNA degradation during extraction and storage, (b) recover cell-free HPV DNA in addition to cell-associated DNA, (c) process a sufficient volume of urine, and (d) use a first-void sample.

In addition, we found that detectable human DNA in urine may not be a good internal control for sample validity. HPV prevalence data that are based on urine samples collected, stored, and/or processed under suboptimal conditions may underestimate infection rates.

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Acknowledgments

This work was supported by the Industrial Research Fund (with the participation (10 %) of GlaxoSmithKline Biologicals) of the University of Antwerp (IOS/SBO 3501/3494).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Centre for the Evaluation of Vaccination, Vaccine & Infectious Disease Institute, University of Antwerp, Universiteitsplein 1, 2610, Antwerpen, Belgium

    A. Vorsters, S. Biesmans, A. Hens, I. De Coster & P. Van Damme

  2. Laboratory of Experimental Hematology, Tumor Immunology Group, Vaccine & Infectious Disease Institute, Antwerp University Hospital, University of Antwerp, Wilrijkstraat 10, 2650, Edegem, Belgium

    J. Van den Bergh

  3. Applied Molecular Biology Research Group, Laboratory of Cell Biology & Histology, University of Antwerp, Groenenborgerlaan 171, 2020, Antwerpen, Belgium

    I. Micalessi & J. Bogers

  4. Laboratory for Microbiology, Vaccine & Infectious Disease Institute, Antwerp University Hospital, University of Antwerp, Wilrijkstraat 10, 2650, Edegem, Belgium

    M. Ieven

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  1. A. Vorsters
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  2. J. Van den Bergh
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  3. I. Micalessi
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  4. S. Biesmans
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  5. J. Bogers
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  6. A. Hens
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  7. I. De Coster
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  8. M. Ieven
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  9. P. Van Damme
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Correspondence to A. Vorsters.

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Vorsters, A., Van den Bergh, J., Micalessi, I. et al. Optimization of HPV DNA detection in urine by improving collection, storage, and extraction. Eur J Clin Microbiol Infect Dis 33, 2005–2014 (2014). https://doi.org/10.1007/s10096-014-2147-2

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  • DOI: https://doi.org/10.1007/s10096-014-2147-2

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