Clinical validation of full HR-HPV genotyping HPV Selfy assay according to the international guidelines for HPV test requirements for cervical cancer screening on clinician-collected and self-collected samples
Authors: Avian Alice , Clemente Nicolò , Mauro Elisabetta , Isidoro Erica , Di Napoli Michela , Dudine Sandra , Del Fabro Anna , Morini Stefano , Perin Tiziana , Giudici Fabiola , Cammisuli Tamara , Foschi Nicola , Mocenigo Marco , Montrone Michele , Modena Chiara , Polenghi Martina , Puzzi Luca , Tomaic Vjekoslav , Valenti Giulio , Sola Riccardo , Zanolla Shivani , Vogrig Enea , Riva Elisabetta , Angeletti Silvia , Ciccozzi Massimo , Castriciano Santina , Pachetti Maria , Petti Matteo , Centonze Sandro , Gerin Daniela , Banks Lawrence , Marini Bruna , Canzonieri Vincenzo , Sopracordevole Francesco , Zanconati Fabrizio , Ippodrino Rudy 
Published: May 2022

Abstract
Background
According to international guidelines, Human Papillomavirus (HPV) DNA tests represent a valid alternative to Pap Test for primary cervical cancer screening, provided that they guarantee balanced clinical sensitivity and specificity for cervical intraepithelial neoplasia grade 2 or more (CIN2+) lesions. The study aimed to assess whether HPV Selfy (Ulisse BioMed – Trieste, Italy), a full-genotyping HPV DNA test that detects and differentiates 14 high-risk HPV (HR-HPV) types, meets the criteria for primary cervical cancer screening described in the international guidelines, on clinician-collected as well as on self-collected samples.

Methods
For each participant woman, consecutively referring to Azienda Sanitaria Universitaria Giuliano Isontina (Trieste, Italy) and CRO—National Cancer Institute (Aviano, Italy) for the cervical cancer screening program, the following samples were tested: (a) a clinician-collected cervical specimen, analyzed with the reference test (Hybrid Capture®2 test, HC2) and HPV Selfy; and (b) a self-collected vaginal sample, analyzed with HPV Selfy. Enrolled women were also asked to fulfill a questionnaire about self-sampling acceptability. As required by guidelines, a non-inferiority test was conducted to compare the clinical performance of the test under evaluation with its reference test.

Results
HPV Selfy clinical sensitivity and specificity resulted non-inferior to those of HC2. By analysis of a total of 889 cervical liquid-based cytology samples from a screening population, of which 98 were from women with CIN2+, HPV Selfy showed relative sensitivity and specificity for CIN2+ of 0.98 and 1.00 respectively (non-inferiority score test: P = 0.01747 and P = 0.00414, respectively); the test reached adequate intra- and inter-laboratory reproducibility. Moreover, we demonstrated that the performance of HPV Selfy on self-collected vaginal samples was non-inferior to the performance obtained on clinician-collected cervical specimen (0.92 relative sensitivity and 0.97 relative specificity). Finally, through HPV Selfy genotyping, we were able to describe HPV types prevalence in the study population.

Conclusions
HPV Selfy fulfills all the requirements of the international Meijer’s guidelines and has been clinically validated for primary cervical cancer screening purposes. Moreover, HPV Selfy has also been validated for self-sampling according to VALHUDES guidelines. Therefore, at date, HPV Selfy is the only full-genotyping test validated both for screening purposes and for self-sampling.
Trial registration ASUGI Trieste n. 16008/2018; CRO Aviano n.17149/2018