JCM Accepts, published online ahead of print on 28 October 2009

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J. Clin. Microbiol. doi:10.1128/JCM.00991-09
Copyright (c) 2009, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Abundance of multiple high-risk HPV infections in cervical cells analyzed by an ultra-sensitive HPV genotyping assay

Markus Schmitt, Bolormaa Dondog, Tim Waterboer, Michael Pawlita^*, Massimo Tommasino, and Tarik Gheit

Department Genome Modifications and Carcinogenesis (F020), Research Program Infection and Cancer, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 242, 69120 Heidelberg, Germany; Infections and Cancer Biology Group, International Agency for Research on Cancer, 150, cours Albert-Thomas, 69372 Lyon, France

^* To whom correspondence should be addressed. Email: m.pawlita{at}dkfz.de.

Polymerase chain reaction (PCR) methods enable the detection of a large variety of human papillomavirus (HPV) genotypes that infect the anogenital tract. However, PCR using consensus or general and, to a lesser extent, broad-spectrum primers may underrepresent the true viral prevelance, especially of multiple infections.

In 735 selected cervical scrapes we compared HPV positivity determined by broad-spectrum BSGP5+/6+-PCR (BS) coupled to an established bead-based Multiplex HPV Genotyping (MPG) assay with that determined by a multiplex type-specific PCR (TS) coupled to a newly-developed MPG assay. While the primers of the BS-PCR are located within the L1 region of the HPV genome, the TS primers target the E7 gene. The overall rate of HPV posititivity for the 19 types included in both assays was 60.9% and 72.2% with BS and TS, and multiple infections were found in 34.8% and 58.0%, respectively. Both HPV detection assays allowed a semi-quantitative detection of HPV types and identified the same dominant HPV type in 66.6% of multiple infections.

In conclusion, the TS-PCR-MPG assay significantly increased the HPV DNA detection rate, the number of multiple infections and demonstrated that the prevalence of low copy HPV infections in the anogenital tract may be strongly underestimated using conventional HPV amplification methods, especially in multiple infections. As a consequence, TS-MPG appears to be highly suited to analyze the significance of multiple infections in the development of cervical cancer and to study the natural history and latency of HPV.