Clinical Validation of the HPV-Risk Assay, a Novel Real-Time PCR Assay for Detection of High-Risk Human Papillomavirus DNA by Targeting the E7 Region

Abstract

Persistent infection with high-risk human papillomavirus (HPV) is the causative agent for cervical cancer (1, 2). Testing for HPV DNA provides better protection against cervical cancer and its precursors, i.e., high-grade cervical intraepithelial neoplasia (CIN), compared to cytology (3-7). For primary cervical cancer screening, it is crucial that the HPV assays that are used are clinically validated to ensure optimal distinction between HPV infections associated with CIN grade 2 or worse (CIN2ϩ) and clinically irrelevant transient HPV infections (8,9).A variety of HPV DNA detection assays are currently considered clinically validated with cervical scraping specimens for cervical cancer screening purposes. Validation has been based on either data from large prospective screening trials (i.e., high-risk HPV Hybrid Capture 2 [HC2] and GP5ϩ/6ϩ PCR) (3, 5, 6, 10) or cross-sectional clinical equivalence analyses according to international guidelines (8, 9) for HPV DNA test requirements (11-13). In addition to clinician-based sampling, HPV self-sampling is an emerging effective strategy for cervical screening. Offering HPV testing on self-collected cervicovaginal specimens reattracts a substantial number of nonattendees into the screening program and effectively detects CIN2ϩ (14, 15). However, standardization of the collection device, HPV test, and sample preparation protocols is important to minimize variations in the CIN2ϩ sensitivity and specificity of HPV self-sampling (reviewed in reference 16). It must be realized that the use of an HPV test that is clinically validated for cervical scraping specimens does not automatically result in high clinical accuracy when it is applied to self-collected specimens (17). Therefore, a separate analysis of the candidate HPV test with self-collected samples, relative to its performance with cervical scraping specimens, is important to ensure suitability for HPV self-sampling. Given the potential variations in target cell yields between different self-samplers (16), such a comparative accuracy analysis ideally should be performed for each selfsampler type, in order to determine the best combination of selfsampler and validated HPV test.Most assays validated for cervical screening purposes use PCRbased assays targeting regions within the HPV E1 or L1 open reading frames (11,13,18). However, malignant progression of cervical lesions is often associated with viral DNA integration into the genome of the host cell (19). Integration often takes place between

Offering Self-Sampling Kits for HPV Testing to Reach Women Who Do Not Attend in the Regular Cervical Cancer Screening Program

“…The aforementioned meta-analysis demonstrated that the sensitivity and specificity of HPV testing are similar on self-as on cliniciantaken samples when validated PCR tests are used but not when signal amplification-based HPV assays are applied (12). The conclusions of this meta-analysis remain unchanged after the addition of recently published studies (12,(14)(15)(16). The pooled relative sensitivity and specificity of 19 studies using HC2 (Qiagen) in self-versus clinician-collected samples were 0.86 (95% CI, 0.82-0.91) and 0.96 (95% CI, 0.93-0.98), respectively (see Fig.…”

Host-cell DNA methylation patterns during high-risk HPV-induced carcinogenesis reveal a heterogeneous nature of cervical pre-cancer

“…HrHPV was detected by GP5+/6+ PCR enzyme immunoassay [56] or HPV-Risk assay (Self-screen B.V.) [57]. This study followed the ethical guidelines of the Institutional Review Board of the VU University Medical Center and University Medical Center Groningen.…”

The prevelance of human papillomavirus (HPV) genotypes detected by PCR in women with normal and abnormal cervico-vaginal cytology

“…The assay uses 16 specific primers amplifying within E6 and E7 regions of the HPV genome and permits the detection of 13 high risk genotypes (16,18,31,33,35,39,45, 51, 52, 56, 58, 59 and 68) and two low-risk genotypes (6 and 11). The presence of DNA of these HPV subtypes were detected by multiplex PCR amplification using type specific primers [14,15].…”