Clinical Validation of a Type-Specific Real-Time Quantitative Human Papillomavirus PCR against the Performance of Hybrid Capture 2 for the Purpose of Cervical Cancer Screening

Abstract

cTo be acceptable for use in cervical cancer screening, a new assay that detects DNA of high-risk human papillomavirus (hrHPV) types must demonstrate high reproducibility and performance not inferior to that of a clinically validated HPV test. In the present study, a real-time quantitative PCR (qPCR) assay targeting the E6 and E7 genes of hrHPV was compared with Hybrid Capture 2 (hc2) in a Belgian cervical cancer screening setting. In women >30 years old, the sensitivity and specificity for intraepithelial neoplasias of grade 2 or worse (93 cases of cervical intraepithelial neoplasias of grade 2 or worse (CIN2؉) and 1,207 cases of no CIN or CIN1) were 93.6% and 95.6%, respectively, and those of hc2 were 83.9% and 94.5%, respectively {relative sensitivity of qPCR/hc2 ‫؍‬ 1.12 [95% confidence interval (CI), 1.01 to 1.23]; relative specificity ‫؍‬ 1.01 [95% CI, 0.99 to 1.03]}. A score test showed that the sensitivity (P < 0.0001) and specificity (P < 0.0001) of the qPCR assay were not inferior to those of hc2 at the required thresholds of 90% and 98%, respectively. The overall agreement of hrHPV positivity between the two runs of the qPCR tests was 98.7% (95% CI, 97.5 to 99.4%), with a kappa value of 0.96 (95% CI, 0.83 to 1.00). The qPCR assay used in this study can be considered a reliable HPV assay that fulfills the clinical validation criteria defined for use in cervical cancer screening.

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].…”

Clinical Evaluation of a GP5+/6+-Based Luminex Assay Having Full High-Risk Human Papillomavirus Genotyping Capability and an Internal Control

“…The Hybrid Capture 2 (HC2; Qiagen, Hilden, Germany) assay (2-4) and GP5ϩ/6ϩ PCR-based enzyme immunoassay (EIA) kit HPV GP HR (EIA; Diassay, Rijswijk, the Netherlands) (5-9) were the first HPV tests to be clinically validated for primary screening on the basis of longitudinal results from large screening studies (10). Testing for high-risk HPV (hrHPV) nucleic acids is also useful for triage of women with equivocal or mildly abnormal cytology for colposcopy and as a test of cure of treatment (10).Several novel HPV tests have been fully or partially validated by showing noninferiority to HC2 or EIA and high reproducibility (11), as has been extensively reviewed by Arbyn et al (10) and reported in additional studies (12)(13)(14). These tests usually detect around 14 hrHPVs in aggregate, but some provide concurrent (partial) genotype-specific information.…”

“…Several novel HPV tests have been fully or partially validated by showing noninferiority to HC2 or EIA and high reproducibility (11), as has been extensively reviewed by Arbyn et al (10) and reported in additional studies (12)(13)(14). These tests usually detect around 14 hrHPVs in aggregate, but some provide concurrent (partial) genotype-specific information.…”

Performance of a Cartridge-Based Assay for Detection of Clinically Significant Human Papillomavirus (HPV) Infection: Lessons from VALGENT (Validation of HPV Genotyping Tests)

“…Intralaboratory testing took place at the AML laboratory (Antwerp, Belgium) and interlaboratory testing took place between AML and the laboratory of the University Hospital of Ghent (Ghent, Belgium). A total of 510 samples collated at the AML laboratory, which had previously been tested with an in-house multiplex real-time PCR (15,16), were assessed. For the validation criteria to be satisfied, the 95% lower confidence bound of both agreements should be Ͼ87%, with a kappa value Ն0.5 (5).…”