Digene hc2 high risk hpv dna test

HPV was detected at the Virology Service of IPO Porto as part of routine procedures using digene HC2 High-Risk HPV DNA Test (QIAGEN, Germany) (HC2). HC2 test detects 13 high-risk HPV (HR-HPV) types: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, and 68. HPV infection was detected in 67.3% of all cervical specimens, with a prevalence of 40.8% (20/49) for normal cytology, 75.0% (21/28) for LSIL, 96.6% (28/29) for HSIL, and 87,5% (7/8) for ICC.

The HC2 solution uses the digene® HC2 High Risk HPV DNA Test® (QIAGEN, Gaithersburg, MD), which is based on signal amplification using RNA probes to target the entire hrHPV genome. Samples were placed in the M1 decapper system where barcode identification, homogenization, decapping/recapping and sample transfer to a bar-coded tube took place. Subsequently, automated sample preparation of cervical cells was performed using the QIAsymphony DSP HPV Media Kit in combination with the QIAsymphony. Resulting sample extracts were tested for hrHPV using the digene® HC2 High-Risk HPV DNA Test® with the Rapid Capture System. Automated amplified chemiluminescent signal detection and results reporting (hrHPV positive: relative light unit per cut off value (RLU/CO) ≥1.0; hrHPV negative RLU/CO < 1.0) was performed using the DML3000 luminometer. All steps were performed according to the manufacturer’s protocols.

The Ventana Benchmark XT platform for ISH (Ventana) was used for HPV detection. Briefly, sections were assayed for HPV DNA by in situ hybridization with INFORM HPV-III Family-16 Probe(B) cocktail for 12 high-risk genotypes, and visualized using the ISH iVIEW PlusDetection Kit (Ventana). The high-risk HPV ISH test was scored as positive if there was any blue reaction product that co-localized with the nuclei of malignant cells.
The digene HC2 High-Risk HPV DNA Test (Qiagen, Germany) was used as a confirmatory assay for HPV detection. The test allows for the qualitative detection of 13 high-risk genotypes. Assays were performed following the manufacturer’s instructions and the chemiluminescent signals were measured in a DML instrument. Samples with processed values ≥1.0 are considered positives.

During the initial screening visit, exfoliative cervical samples were taken with a Cervex-Brush (Rovers Medical Devices, Oss, Netherlands) and used for conventional cytology, which was classified according to the Bethesda System as negative, atypical squamous cells of undetermined significance (ASC-US), low-grade squamous intraepithelial lesion (LSIL), and high-grade squamous intraepithelial lesion (HSIL). Immediately after cytology, the residual cells remaining on the brush were rinsed into vials containing 20 mL of preservative ThinPrep PreservCyt medium (Hologic, Marlborough, MA, USA) for HPV testing. HPV testing was performed according to manufacturer's instructions at independent laboratories using the digene HC2 High-Risk HPV DNA Test (QIAGEN, Germantown, MD, USA) or COBAS 4800 HPV Test (Roche Diagnostics, Mannheim, Germany). Between December, 2012, and May, 2013, HPV testing was done in Colombia on 872 samples stored on the digene Specimen Transport Medium.