Cervical swabs and semen samples were tested for the presence of HPV DNA using the cobas® HPV Test (Roche Diagnostics GmBH, Mannheim, Germany) according to the manufacturer’s recommendations for cervical swabs management [12] and using the PapilloCheck® test (Greiner Bio-One, Frickenhausen, Germany) [13] as described previously [5]. DNA from penile swabs was isolated by QIAamp® DNA Micro kit (Qiagen, Hilden, Germany) and eluted with 40 μL of diethyl pyrocarbonate (DEPC) treated water. Isolated DNA was then tested using PapilloCheck® [13]. Samples with HPV detection failure were repeated three times. HPV detection repeatedly failed in 71 penile swab samples and 1 semen sample due to insufficient amount of genetic material. Participants with failed samples were excluded from the study.

The cobas® HPV Test and the PapilloCheck® test gave discordant results for 8 out of 195 cervical swabs and 4 out of 195 semen samples included in the study. These 12 samples were additionally tested using the LMNX Genotyping Kit HPV GP (Diassay, Rijswijk, The Netherlands) [14] as described previously [5]. The final HPV result for a given cervical swab or semen sample was determined by concordant results of at least two HPV detection methods, as described previously [6]. The HPV result for a given penile swab was based only on the PapilloCheck® test (Fig. 2).

Methods used for HPV detection in different sample types

We examined any-type concordance (both partners are HPV positive), same-type concordance (both partners are HPV positive for 1 or more mutual HPV types), and type-specific concordance for vaccine-targeted genotypes HPV16, 18, 31, 33, 45, 52, 58, 6, and 11. For this type of analysis, men was considered as HPV positive when either the penile swab or the semen sample was HPV positive.

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