**5. Conclusions**

Having in mind that any technical innovation is usually related to new perspectives for both clinical and technical development over time, the results obtained sugges<sup>t</sup> that the temporal distribution of HPV genotypes might be influenced by the diagnostic methods that are performed.

This study is not a direct comparison of two diagnostic methods because it has a sequential design. In summary, with univariate analysis significant di fferences were observed in the prevalence rates of HPV-45, 68, 40, 42, and 43. The lowest prevalence for HPV-45 was observed in the Magna Pure-Real Time PCR group, while HPV-68, 40, 42, and 43 were less observed in the Qiagen-Real Time PCR group.

With logistic regression analysis, the introduction of the real-time assay was associated with an increase in HPV-42. Furthermore, the logistic regression analysis did not reveal the presence of underlying trends. It is possible that the di fference in the HPV-42 distribution detected after the switch to the real-time test could be related to the di fferent sensitivities of the two diagnostic assays or to the presence of cross-reactions.

An important limitation of this kind of study is the potential confounding e ffect of some variables (secular trends, changes in population composition, or behavioral risks). Unfortunately, these data were not available for the analysis. However, despite the increase in number of subjects tested, the stability of HPV prevalence over time suggests that the population composition and the behavioral variables did not likely change during the observation period.

Therefore, this parameter should be taken into the consideration when a multivariate statistical model is represented, and further clinical studies could increase knowledge on the prevalence of HPV based on the use of di fferent methods.

**Author Contributions:** Conceptualization, R.D.P.; Data curation, L.R., A.A. and L.S.; Investigation, R.D.P., L.R., G.A., R.M. and A.A.; Methodology, R.D.P., L.R., G.A., R.M., D.D.C. and L.S.; Project administration, R.D.P. and L.S.; Supervision, R.D.P. and L.S.; Validation, R.D.P., L.R. and D.D.C.; Writing—original draft, A.A.; Writing—review and editing, L.S.

**Funding:** This study did not receive funding.

**Acknowledgments:** We are extremely thankful to Christopher Williams, University of Foggia, for the text revision.

**Conflicts of Interest:** The authors declare no conflict of interest.
