Clinical Validation of the Vitro HPV Screening Assay for Its Use in Primary Cervical Cancer Screening.

Many scientific societies have issued guidelines to introduce population-based cervical cancer screening with HPV testing. The Vitro HPV Screening assay is a fully automatic multiplex real-time PCR test targeting the L1 GP5+/GP6+ region of HPV genome. The assay detects 14 high risk (HR) HPV genotypes, identifying individual HPV16 and HPV18 genotypes, and the HPV-positive samples for the other 12 HR HPV types are subsequently genotyped with the HPV Direct Flow Chip test. Following international guidelines, the aim of this study was to validate the clinical accuracy of the Vitro HPV Screening test on ThinPrep-collected samples for its use as primary cervical cancer screening, using as comparator the validated cobas® 4800 HPV test. The non-inferiority analysis showed that the clinical sensitivity and specificity of the Vitro HPV Screening assay for a diagnosis of cervical intraepithelial neoplasia of grade 2 or worse (CIN2+) were not inferior to those of cobas® 4800 HPV (p = 0.0049 and p < 0.001 respectively). The assay has demonstrated a high intra- and inter-laboratory reproducibility, also among the individual genotypes. The Vitro HPV Screening assay is valid for cervical cancer screening and it provides genotyping information on HPV-positive samples without further sample processing in a fully automated workflow.

Cervista HPV HR test for cervical cancer screening: a comparative study in the Catalonian population.

CONTEXT: Almost all cervical cancers are related to the human papillomavirus (HPV). Future strategies for cervical cancer screening will be based on HPV detection. The Hybrid Capture 2 (HC2) test is currently the most widely used method to screen for HPV. OBJECTIVE: To test the performance of the Cervista HPV HR test for cervical screening. DESIGN: We examined 875 cervical samples by HC2 and Cervista. Of these, 64 were high-grade cervical intraepithelial neoplasia (CIN 2+) cases and were used to test the sensitivity of the assay. The remaining 811 were non-CIN 2+ cases, which were used to compare specificity. The noninferiority score test was used, with at least 0.90 for sensitivity and 0.98 for specificity and with a κ value of 0.7. RESULTS: Sensitivity and specificity were, respectively, 100% and 86.4% for the HC2 test, and 98.4% and 85.2% for the Cervista test. The agreement between the two assays was 91.7% (802 of 875; κ = 0.743; 95% confidence interval, 0.688-0.798). The noninferiority score test (relative sensitivity of 90%, T = 2.85, P = .002; and relative specificity of 98%, T = 2.75, P = .003) demonstrated that the Cervista results were not inferior to those of the HC2 test. Intralaboratory and interlaboratory reproducibility was determined by evaluating 513 and 507 samples, respectively. These reproducibilities showed κ values of 0.886 (95% confidence interval, 0.845-0.927) and 0.907 (95% confidence interval, 0.886-0.948), respectively. CONCLUSIONS: Our results demonstrate that the Cervista HPV HR test shows the same specificity as the HC2 assay. We therefore conclude that the Cervista HPV HR test is suitable for cervical cancer screening purposes.

Human papillomavirus detection and p16INK4a expression in cervical lesions: a comparative study.

p16(INK4a) expression in dysplastic cervical lesions is related to high-risk human papillomavirus (HR-HPV) infection. The immunohistochemical expression of this protein in these lesions allows an increase in diagnostic reproducibility in biopsies and the introduction of prognostic factors in low-grade lesions. Here, we studied the immunohistochemical expression of p16 in 86 dysplastic cervical lesions, 54 cervical intraepithelial neoplasms-grade 1 (CIN-I), 23 CIN-II, and 9 CIN-III. In addition, we performed HPV detection and genotyping. We detected HR-HPV in 19/54 CIN-I, 21/23 CIN-II and 9/9 CIN-III cases. p16(INK4a) immunoreactivity was observed in 7/19 CIN-I HR-HPV-positive, 17/21 CIN-II HR-HPV-positive and all CIN-III cases. Immunoreactivity for p16(INK4a) was found in 7/54 CIN-I and in 17/23 CIN-II cases. In the follow-up, we detected 3 p16-positive high-grade squamous epithelial lesions (CIN-II and CIN-III) in the CIN-I/p16-negative group and 5 p16-positive high-grade squamous epithelial lesions cases in the CIN-II/p16-negative group. We conclude that p16 negativity in CIN-I and CIN-II biopsies does not always imply regression of the lesion and that the diagnosis of CIN-II should not be based solely on p16 results.

HPV testing by cobas HPV test in a population from Catalonia.

BACKGROUND: HPV testing in cervical cancer screening has been proposed as an alternative or complementary to cytology in women older than 30 years. However, adequate clinical sensitivity and specificity are crucial for a new test to be implemented. Hybrid Capture 2 (HC2) has proved good clinical performance in selecting women at risk for high-grade intraepithelial lesions with a high sensitivity and specificity. cobas HPV Test has been recently launched and its performance in different clinical settings needs to be determined. OBJECTIVES: The aim of this study was to evaluate the cobas HPV Test for the detection of cervical HPV infection in a population of women in Catalonia (Spain) using HC2 as a reference. MATERIALS AND METHODS: Cervical liquid cytology samples from 958 women have been studied. Sensitivity was analyzed in 60 samples from patients with a high-grade intraepithelial lesion (≥ CIN2) on histology and specificity was determined in 898 samples from women with no ≥ CIN2. All cases had HC2 and cobas HPV Test performed. Statistical analyses of sensitivity, specificity and comparison between HC2 and cobas HPV Test by a non-inferiority test were applied. RESULTS: Sensitivity of HC2 and cobas HPV Test for detecting ≥ CIN2 proved identical (98.3%) while specificity was 85.3% and 86.2% respectively. The non-inferiority test demonstrated that cobas HPV Test surpassed 90% sensitivity and 98% specificity of HC2. CONCLUSION: The cobas HPV Test results fulfilled sensitivity and specificity requirements for HPV based cervical cancer screening and for the triage of minor cytological abnormalities, allowing its introduction in clinical settings.

Estudio comparativo de la determinación de HPV entre HC2 y Cervista™ / Comparative study of HC2 and Cervista™ for the detection of HPV

Antecedentes. La determinación del virus del papiloma humano (VPH) en muestras ginecológicas es en la actualidad un elemento clave para la toma de decisiones en patología cervical. El método más contrastado clínicamente es la captura de híbridos de segunda generación (HC2). Métodos. Estudio comparativo de los resultados obtenidos, mediante HC2 y Cervista™, para la detección de VPH de alto riesgo (VPH-AR) en muestras ginecológicas. Resultado. La comparación de resultados muestra una concordancia del 96,4%, con una sensibilidad de Cervista™ para CIN2+ del 98,3% y una especificidad del 98,4%. Conclusiones. Creemos que Cervista™ es un buen método para la detección de VPH-AR, aplicable tanto en el cribado como en la clínica(AU)

Introduction. HPV detection in gynaecological samples currently provides key data in cervical pathology for diagnostic and management decisions. At present HC2 is the most clinically proven method for this purpose. Methods. Comparative study of HPV-HR detection between HC2 and Cervista™. Results. The concordance between the two methods is 96.4%. The sensitivity of Cervista™ for CIN2+ is 98.3% and specificity exceeds 98.4%. Conclusions. We conclude that Cervista™ is an appropriate method for the detection of HPV-HR in gynecological samples for screening and clinical purposes(AU)

Detection of HPV by in situ hybridization in thin-layer (ThinPrep) cervicovaginal samples.

We have studied an automated in situ hybridization (ISH) method as a possible alternative approach for detecting high-risk human papillomavirus (HPV) in monolayer (ThinPrep) cervico-vaginal samples, comparing the results with those obtained by polymerase chain reaction (PCR) using consensus primers and studying the relationship between the ISH staining pattern and the viral integration in HPV 16-positive cases. Eighty atypical squamous cells of undetermined significance (ASCUS) and low-grade squamous intraepithelial lesion (LSIL) cases were used for our purposes. The patients were monitored through periodic cytologies. ISH with was performed with an automated Ventana System, analysis by PCR was performed with consensus primers and integration of HPV16 was performed by realtime PCR analyzing E2 and E6 genes. Additionally, 27 HSIL cases were also studied to observe the ISH staining patterns. HPV infection was detected by ISH in 21.7% of the ASCUS cases and 55.8% of the LSIL cases. Two distinct staining patterns were observed: multipunctated (MP) and diffuse (DI). In some cases, a mixed pattern (MP + DI) was observed and these cases were considered as MP. The MP pattern increased with the degree of lesion and seemed to have a prognostic value in ASCUS/LSIL cases. The lesion in MP pattern cases persisted throughout the entire study in 77% of cases, whereas in cases with a DI staining pattern, only 41% of them showed persistence of the lesion (p <0.001). No correlation was found between HPV integration and the ISH staining pattern. Given the lower sensitivity and negative predictive value of ISH and its incapacity to demonstrate the integration of high-risk HPV in ASCUS and LSIL cases using liquid-based cytology, we do not recommend this technique for the triage of ASCUS and LSIL cases.