Cervical cancer screening based on Pap Test represents one of the most successful public health interventions in the last 60 years, having led to the decrease of both cervical cancer incidence and mortality [
1]. Recently, randomized control trials, using either of these two high-risk human papillomavirus (hrHPV) DNA testing methods: Hybrid Capture 2 (HC2, QIAGEN, Germany) or GP5+/GP6+ PCR-EIA, have led to recognize them as clinically validated alternatives for use in primary cervical cancer screening, providing 60–70% greater protection compared with cytology [
2,
3]. In general, when used for primary screening, any hrHPV DNA test has to guarantee a balanced clinical sensitivity and specificity in order to allow effective detection of cervical intraepithelial neoplasia grade 2 (CIN2) or more severe lesions (>CIN2) and minimize follow up procedures on HPV test-positive women without clinically meaningful disease (<CIN2) [
4,
5]. Furthermore, high intra- and inter-laboratory reproducibility is required to ensure reliable performance of the test in clinical practice. In order to guarantee these specification, recent guidelines [
6] have proposed a clinical validation strategy based on the comparison of the performance of the assay under evaluation with that of an already clinically validated reference hrHPV test, on samples originating from a population-based screening cohort. In accordance with the above mentioned guidelines, a retrospective study was set up aiming to assess the clinical specificity and sensitivity of REALQUALITY RQ-HPV Screen (AB ANALITICA, Italy) in association with the automated platform GENEQUALITY X120 (AB ANALITICA, Italy). This test, based on real time PCR, targets the E6-E7 region of 14 hrHPV genotypes. The standard comparator test chosen to allow expression of performance in relative terms was HC2, which, besides providing adequate clinical sensitivity and specificity for screening purposes [
5‐
7], is also FDA approved. The reproducibility of the test under evaluation was assessed as well.