Clinical Validation of the cobas 4800 HPV Test for Cervical Screening Purposes

Abstract

This study shows that the clinical performance and reproducibility of the cobas 4800 HPV test for high-risk human papillomavirus (HPV) detection fulfill the criteria as formulated in international guidelines of HPV test requirements for cervical screening purposes. Accordingly, the cobas 4800 HPV test can be considered clinically validated for cervical screening.A critical feature for high-risk human papillomavirus (hrHPV) tests used for cervical screening is their clinical accuracy for detection of high-grade cervical intraepithelial neoplasia (CIN) and cervical cancer (CIN2ϩ). For primary screening, an hrHPV test should have a balanced clinical sensitivity and specificity to allow effective detection of CIN2ϩ and minimize follow-up procedures of HPV test-positive women without CIN2ϩ. Furthermore, high intra-and interlaboratory reproducibility is required to ensure reliable performance of the test in clinical practice. Both the high-risk HPV Hybrid Capture 2 method (HC2) and GP5ϩ/6ϩ-PCR-enzyme immunoassay (EIA) fulfill these specifications and are considered clinically validated for screening purposes (4). In order to facilitate the validation procedure of any novel, candidate hrHPV assay without the necessity of performing large, prospective screening trials, specific criteria have recently been outlined by an international consortium based on clinical equivalence analysis (3, 4). Here, we evaluated the recently FDA-approved cobas 4800 HPV test (1) according to this validation strategy. The cobas 4800 HPV test features automated sample preparation combined with real-time PCR technology to detect 14 hrHPV genotypes. PCR amplification and detection occur in a single tube, where probes with four different reporter dyes track the different targets in the multiplex reaction: (i) HPV16, (ii) HPV18, (iii) 12 hrHPV types (i.e., HPV31, as a pool, and (iv) ␤-globin as the control for extraction and amplification adequacy.The clinical performance of the cobas 4800 HPV test was assessed relative to HC2, which detects 13 hrHPV types (i.e., HPV16,, in a cohort of screening participants originally screened by cytology and HC2 cotesting (intervention group of the VUSA-Screen trial) (5, 6). A detailed description of the VUSA-Screen trial, including the referral policy and follow-up procedure, has been published previously (6). Informed consent was obtained from all study participants, and this study followed the local ethical guidelines of the medical center.A total of 860 archived cervical scrapings were selected comprising (i) a set for clinical sensitivity analysis of 60 representative scrapes from women (median age of 35 [range, 29 to 60] years) who had histologically confirmed CIN2ϩ (i.e., 23 with CIN2, 33 with CIN3, 1 with adenocarcinoma, and 3 with squamous cell carcinoma) diagnosed within a median follow-up time of 3.3 (range, 0 to 32.6) months and detected through either HC2 or cytology or both (hereafter referred to as cases) and (ii) a set for clinical specificity analysis of 800 representative scrapes from women (m...

Evaluation of HPV Molecular Tests in Primary Screening for Cervical Cancer in Brazil

“…before any processing. Hybrid Capture 2 and cobas 4800 HPV test showed very similar performances (sensitivity and specificity for CIN3+ of 84% and 89% for both tests respectively) corroborating findings from other comparative evaluations [26]. Cobas 4800 offers the advantage of genotyping HPV 16 and 18 in the screening test and also contains a control for sample adequacy (beta-globin), being a fully automated platform.…”

Clinical Validation of the BD Onclarity? HPV Assay Using a Non-Inferiority Test

“…both showing a percentage of agreement with a lower confidence bound not less than 87% (kappa value of at least 0.5)) [4]. Recently a number of different molecular HPV test methods have been subjected to this test criterion and it has become the de facto standard by which new tests are assessed [5][6][7].…”

“…Since the introduction of the HC2 assay, the majority of HPV assays have been developed using polymerase chain reaction (PCR) technology that utilizes consensus PCR primers targeting the L1 region of the virus genome [6][7][8]. While L1-based amplification assays have been shown to have clinical performance comparable to clinically validated assays such as HC2 [9,10], there are certain technical limitations to the use of a consensus primer approach, namely, inconsistency in detecting low copy number infections within mixed infections [11][12][13], and the potential risk of missing late stage cancers due to deletion of the L1 target region [14,15].…”

Comparison of Seegene Anyplex II HPV28 with the PGMY-CHUV Assay for Human Papillomavirus Genotyping

“…Assays that have been clinically validated identify high-risk genotypes as a whole (the hybrid capture II [HCII] assay) or distinguish HPV16 or HPV18 from the other high-risk types as a group (HPV31, -33, -35, -39, -45, -51, -52, -56, -58, -59, -66, and -68) (Abbott RealTime high-risk HPV and Roche Cobas 4800 HPV tests). The Abbott and Roche tests have been validated against HCII using cervical intraepithelial neoplasia (CIN) grade 2 or higher as endpoints (6)(7)(8). They were also found in these studies to be suitable for primary cervical cancer screening according to published guidelines (9).…”