Anal samples were obtained by introducing a cytobrush (Eurogine S

Anal samples were obtained by introducing a cytobrush (Eurogine SL, Sant Boi del Llobregat, Spain) into the anal canal to a depth of 3 cm and gently rotating for 30–45 seconds. The cytobrush was included and shaken into a solution of 20 mL of PreservCyt/ThinPrep Pap test (Cytyc Iberia SL, Barcelona, Spain) for 30 seconds and the solution was stored at −20°C until analysis. DNA was extracted from cell suspensions (in ThinPrep Pap solution) using the Qiamp Viral DNA kit (Qiagen, Hilden, Germany). HPV detection and typing were PD0325901 performed on all samples using a commercial In Vitro Diagnostics with the CE mark certification (IVD-CE) marked assay: the Multiplex Fluorescent-PCR

Kit for Human Papilloma Virus Genotyping (F-HPV typing™; Molgentix SL, Barcelona, Spain) in accordance with the manufacturer’s instructions [20]. The kit allows the detection of 13 HR HPV genotypes: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68; and the two most frequent LR HPV genotypes: 6 and 11. A human short tandem repeat (STR) sequence

included in the same multiplex reaction was amplified as an internal control for DNA integrity and absence of PCR inhibitors. Products selleck compound were analysed by capillary electrophoresis on an ABI 3130 XL genetic analyser and using GeneMapper 4.0 software (Applied Biosystems, City Foster, CA). Each PCR run included HPV-positive and -negative controls. Particular care was taken to prevent

carry-over contamination by separating pre- and post-PCR areas in the laboratory. Cytological changes were classified according to the Bethesda System: atypical squamous cells of uncertain significance (ASCUS) and low- and high-grade squamous intraepithelial lesions (LSILs and HSILs, respectively). Samples were independently Wilson disease protein assessed by two expert cytopathologists. Baseline characteristics were summarized with standard descriptive statistics and a descriptive analysis was carried out. The prevalences of overall anal HPV infection, HPV type-specific infection, and cytological diagnoses were estimated. Differences between groups were evaluated using the χ2 test for qualitative variables, and odds ratios (ORs) and their 95% confidence intervals (95% CIs) were calculated. Bivariate and multivariate logistic (for HPV infection) and nominal (for cytological changes) regression models were used where appropriate. For the multivariate analyses, factors with P < 0.2 in the bivariate model or clinical relevance were used to assess interactions in the multivariate regression model. A P-value ≤ 0.05 was considered statistically significant. Data analysis was carried out using spss version 15.0 statistical software (SPSS, Chicago, IL). The CARH·MEN cohort comprised 733 patients recruited from January 2005 to May 2009.

Comments are closed.