Clinical validation of the HPVIR high-risk HPV test on cervical samples according to the international guidelines for human papillomavirus DNA test requirements for cervical cancer screening

Abstract

Background The indicating FTA card is a dry medium used for collection of cervical samples. HPVIR is a multiplex real-time PCR test that detects 12 high-risk human papillomavirus types (hrHPV) and provides single genotype information for HPV16, − 31, − 35, − 39, − 51, − 56, and − 59 and pooled type information for HPV18/45 and HPV33/52/58. The aim of this study was to evaluate whether a strategy with cervical samples collected on the FTA card and subsequently analysed with the HPVIR test complies with the criteria of the international guidelines for a clinically validated method for cervical screening. Methods We performed a non-inferiority test comparing the clinical sensitivity and specificity of the candidate test (FTA card and HPVIR) with a clinically validated reference test (Cobas ® HPV test) based on liquid-based cytology (LBC) samples. Two clinical samples (LBC and FTA) were collected from 896 participants in population-based screening. For evaluation of the specificity we used 799 women without ≥ CIN2, and for clinical sensitivity we used 67 women with histologically confirmed ≥ CIN2. The reproducibility was studied by performing inter- and intra-laboratory tests of 558 additional clinical samples. Results The clinical sensitivity and specificity for samples collected on the FTA card and analysed using the HPVIR test were non-inferior to samples analysed with the Cobas® HPV test based on LBC samples (non-inferiority test score, p = 1.0 × 10 − 2 and p = 1.89 × 10 − 9 , respectively). Adequate agreement of > 87% was seen in both the intra- and inter-laboratory comparisons. Conclusions Samples collected on the indicating FTA card and analysed with HPVIR test fulfil the requirements of the international guidelines and can therefore be used in primary cervical cancer screening.

Acceptability of self- collection for human papillomavirus detection in the Eastern Cape, South Africa

“…The eluted DNA from specimens on FTA cards was tested for HR-HPV using the clinically validated hpVIR real-time PCR assay [29]. hpVIR assay detects 12 HR-HPV genotypes (HPV16, 18,31,33,35,39,45,51,52, 56, 58 and 59) and the analysis was performed as previously described [30].…”

Incidence of HPV and HPV related dysplasia in elderly women in Sweden

“…Most of the HPV tests were performed using a multiplex real-time PCR assay (hpViR), which detects the following high-risk HPV types 16,18,31,33,35,39,45,51,52,56,58 and 59 (18 and 45 are detected together, and 33,52 and 58 as one group) [18]. A cervical sample was collected using a cytobrush.…”

HPV viral load in self-collected vaginal fluid samples as predictor for presence of cervical intraepithelial neoplasia

“…Since the relationship between hrHPV viral load and CIN2+ risk differs between hrHPV types, an HPV test is required that is able to identify individual or specific groups of hrHPVs. Also, the test must enable quantification of hrHPV copy number and titer, such as the clinically validated hpVir test [26]. The most commonly used HPV tests in screening program (Hybrid Capture 2, Cobas HPV) only distinguish HPV16 and 18 from other hrHPV types, and do not report on viral copy number.…”

“…HPV testing was performed using the clinically validated, real-time PCR-based, hpVIR test [25, 26]. This test detects and quantifies HPV16, 31, 35, 39, 51, 56 and 59 as individual genotypes, HPV18 and 45 in one group and HPV33, 52 and 58 as a second group.…”