High performance of a new PCR-based urine assay for HPV-DNA detection and genotyping.

Human papillomavirus (HPV) testing has been proposed as a means of replacing or supporting conventional cervical screening (Pap test). However, both methods require the collection of cervical samples. Urine sample is easier and more acceptable to collect and could be helpful in facilitating cervical cancer screening. The aim of this study was to evaluate the sensitivity and specificity of urine testing compared to conventional cervical smear testing using a PCR-based method with a new, designed specifically primer set. Paired cervical and first voided urine samples collected from 107 women infected with HIV were subjected to HPV-DNA detection and genotyping using a PCR-based assay and a restriction fragment length polymorphism method. Sensitivity, specificity, Positive Predictive Value (PPV), and Negative Predictive Value (NPV) were calculated using the McNemar's test for differences. Concordance between tests was assessed using the Cohen's unweighted Kappa (k). HPV DNA was detected in 64.5% (95% CI: 55.1-73.1%) of both cytobrush and urine samples. High concordance rates of HPV-DNA detection (k = 0.96; 95% CI: 0.90-1.0) and of high risk-clade and low-risk genotyping in paired samples (k = 0.80; 95% CI: 0.67-0.92 and k = 0.74; 95% CI: 0.60-0.88, respectively) were observed. HPV-DNA detection in urine versus cervix testing revealed a sensitivity of 98.6% (95% CI: 93.1-99.9%) and a specificity of 97.4% (95% CI: 87.7-99.9%), with a very high NPV (97.4%; 95% CI: 87.7-99.9%). The PCR-based assay utilized in this study proved highly sensitive and specific for HPV-DNA detection and genotyping in urine samples. These data suggest that a urine-based assay would be a suitable and effective tool for epidemiological surveillance and, most of all, screening programs.

Combined surgery and cidofovir is an effective treatment for genital warts in HIV-infected patients.

OBJECTIVE: To evaluate the efficacy of surgical-cidofovir (SCT), surgical (ST) and cidofovir (CT) treatment of genital warts in HIV-infected patients. DESIGN: Open randomized prospective pilot study. SETTING: Outpatients attending the sexually transmitted disease service of the II Dept of Infectious Diseases, L Sacco Hospital, Milan-Italy. PATIENTS: Consenting HIV-positive patients with anal-genital warts recruited from January 2000 to March 2001. INTERVENTIONS: Three treatment arms: surgical excision by electrocautery, topical 1% cidofovir-gel (5 days per week, maximum 6 weeks) and electrocautery-cidofovir treatment with 1% cidofovir-gel applied within 1 month of surgical treatment (5 days per week for 2 weeks). MAIN OUTCOME MEASURES: Rate of wart clearance and time and rate of relapses within a 6-month follow-up period. RESULTS: Complete response was achieved in 93.1% of 29 patients treated by ST, 76.2% of 26 treated by CT and in 100% of 19 patients treated by SCT (P = 0.0033). The relapse rate in 49 patients followed-up was 73.68% in ST, 35.29% in CT and 27.27% in SCT patients (P = 0.018). Median time to relapses in ST patients was 66 days (Kaplan-Meyer, P = 0.0012). Human papillomavirus DNA was cleared in 52.63% of 19 patients evaluated. The rate of clearance of high risk and low risk genotypes was 0% and 57.14% 25% and 50% 100% and 71.42% in ST, CT and SCT patients, respectively. CONCLUSIONS: A combination of surgical and medical treatment was most effective in clearing lesions completely and in reducing the relapse rate. Human papillomavirus DNA clearance can be attributed to the antiviral effect of cidofovir and could explain the low relapse rate observed. Larger studies are required to determine the most appropriate medical treatment for viral eradication after surgery.

Sexual behaviour reported by a sample of Italian MSM before and after HIV diagnosis.

In 2006 we conducted a cross-sectional study involving hospital clinical centres in five Italian cities to compare the sexual behaviour of HIV-positive MSM (men who have sex with men) before and after the diagnosis of HIV infection. Each centre was asked to enrol 30 HIV-positive persons aged ≥ 18 years. The questionnaire was administered to 143 MSM on average 9 years after HIV diagnosis. After diagnosis there was a decrease in the number of sexual partners: the percentage of persons who reported having had more than 2 partners decreased from 95.8% before diagnosis to 76.2% after diagnosis. After diagnosis, there was a significant decrease in the percentage of persons who had never (or not always) used a condom with their stable partner for anal sex from 69.2% before diagnosis to 26.6% after diagnosis and for oral-genital sex from 74.8% before diagnosis to 51.7% after diagnosis. Though at-risk behaviour seems to decrease after diagnosis, seropositive MSM continue to engage in at-risk practices: one fourth of them did not use a condom during sexually transmitted infections (STI) episodes, 12.5% of the participants had had sex for money, and 8.4% had paid for sex. The study shows that our sample of Italian HIV-positive MSM, though aware of being infected, engage in sexual behaviours that could sustain transmission of HIV and other STIs. The results could constitute the first step in implementing national prevention programs for persons living with HIV.

Human papillomavirus genotypes and phylogenetic analysis of HPV-16 variants in HIV-1 infected subjects in Italy.

A cross-sectional study was carried out to improve the state of evidence regarding the spectrum of HPV types and HPV-16 LCR variants circulating among men and women infected with HIV-1 in Italy. This study, conducted in 518 HIV-positive subjects (346 males and 172 females), showed a high prevalence of HPV anal infections (88.7%) in men and of cervical infections (65.1%) in women. A wide spectrum of HPV genotypes has been observed, as both single and multiple infections. Low-risk HPV types 6, 11 and 61 were frequently detected. HPV-16 was the prevalent high-risk type. Fourteen different HPV-16 LCR variants were found. Ten belonged to the European lineage (78.7% were detected in Italian subjects and 21.3% in foreign-born, all homo/bisexual men), two to the Asiatic lineage and two to the African-2 lineage. This study underlines the great genotypic heterogeneity characterizing anal and cervical HPV infections and the marked polymorphism of the predominant HPV-16 in this high-risk population in Italy.

Anal HPV genotypes and related displasic lesions in Italian and foreign born high-risk males.

Anal intraepithelial neoplasia and anal cancer are closely related to infection from high-risk Human Papilloma Virus (HPV) genotypes. Since HPVs involved in disease progression are reported to vary by geographical regions, this study focuses on HPV genotypes spectrum in 289 males attending a Sexual Transmitted Diseases (STD) unit according to their nationality. Anal cytology, Digene Hybrid Capture Assay (HC2) and HPV genotyping were evaluated in 226 Italian (IT) and 63 foreign born (FB) subjects, recruited between January 2003 and December 2006. FB people were younger (median 32y-IQR 27-35 vs 36y-IQR 31-43, respectively; Mann-Whitney test p<0.0001) and had a higher rate of abnormal results (>or=atypical squamous cells of undetermined significance (ASCUS)) on anal cytology (95.0% vs 84.04%) (p=0.032; OR 3.61; 95% CI 1.04-1.23). HPV-16 is by far the most common genotype found in anal cytological samples independently from nationality while differences in distribution of other HPV genotypes were observed. The probability of infection from high-risk HPVs was higher in FB (OR 1.69; 95% CI 1.07-2.68) and is due to a higher rate of HPV-58 (OR 4.98; 95% CI 2.06-12.04), to a lower rate of HPV-11 (OR 0.35; 95% CI 0.16-0.77), to the presence of other high-risk genotypes (HPV-45, HPV-66, HPV-69). Multiple infections rate was high and comparable between IT and FB people. The relative contribution of each HPV genotype in the development of pre-neoplastic disease to an early age in the FB group cannot be argued by this study and more extensive epidemiological evaluations are needed to define the influence of each genotype and the association with the most prevalent high-risk HPVs on cytological intraepithelial lesions development.

Changes in at-risk behavior for HIV infection among HIV-positive persons in Italy.

Many HIV-positive persons reportedly continue to engage in at-risk behavior. We compared the sexual and drug-using practices of HIV-positive persons before and after the diagnosis of HIV infection to determine whether their behavior had changed. To this end, in 2006, we conducted a cross-sectional study involving clinical centers in five Italian cities. Each center was asked to enroll 100 persons aged 18 years or older who had a diagnosis of HIV infection that dated back at least 2 years. Data were collected with a specifically designed questionnaire, administered during a structured interview. The McNemar chi2 test was used to compare the data before and after the diagnosis. A total of 497 persons participated (65.5% males; median age of 40 years; age range, 34-45 years). The most common exposure categories were: heterosexual contact (43.4%), homosexual contact (27.2%), and injecting drug use (20.6%). Although the percentage of drug users significantly decreased after diagnosis, 32.4% of injectors continued to use drugs, and approximately half of them exchanged syringes. Regarding sexual behavior, after diagnosis there was a significant decrease in the number of sexual partners and in stable relationships and an increase in condom use, both for persons with stable partners and those with occasional partners, although the percentage varied according to the specific sexual practice. These results indicate that though at-risk behavior seems to decrease after the diagnosis of HIV infection, seropositive persons continue to engage in at-risk practices, indicating the need for interventions specifically geared toward HIV-positive persons.

Percutaneous treatment of femoro-popliteal aneurysms with covered stents.

PURPOSE: To assess the efficacy of the percutaneous treatment of femoro-popliteal aneurysms with covered stents. MATERIALS AND METHODS: Ten femoro-popliteal (1 femoral, 6 femoro-popliteal, 3 popliteal) aneurysms with diameters ranging from 2.1 to 6 cm (mean 3.6 cm) and lengths of 2.2-9 cm (mean 5.8 cm) were treated between September 1998 and December 2001. The patients were 8 men and 2 women aged between 33 and 73 years (mean age 65.4). Self-expanding covered stents (Wallgraft) were employed in all cases: the flexible and elastic stents were implanted percutaneously, under local anesthesia, using 9-10 F introducer sheaths. Color-Doppler ultrasound confirmed the clinical diagnosis. In all cases spiral CT angiography was performed to better evaluate the diameters and lengths of the aneurysms and the size of the arteries above and below the aneurysms. The patients were followed up with color-Doppler US and only occasionally with CT angiography. RESULTS: Stent implantation was successful in all patients; there were no complications. As regards follow up, 6 patients did not show any stent-graft occlusions. One patient, who died 45 days later of causes unrelated to the procedure, was not taken into account. As for the remaining five patients, color-Doppler US showed primary patency of the stents and exclusion of aneurysms at 24 months (2/5), 12 months (1/5), and 6 months (2/5). There were 4 cases of stent-graft occlusion within the first month after placement (in 3/4 cases the caudal end of the aneurysm was located in the median segment of the popliteal artery), re-canalized by local thrombolysis and, in two cases, by PTA aimed at removing the stenoses caused by intimal hyperplasia: one case was patent at 36 months, 3 cases re-occluded and were converted to surgical by-pass (1 at 14 months, 2 at 18 months). Therefore in our series (mean follow-up: 18 months) primary patency was 55.5% and secondary patency was 66.6%. CONCLUSIONS: Percutaneous placement of a covered stent seems to provide an alternative to surgery for aneurysms with limited length and caudal end located not beyond the proximal tract of the popliteal artery. A good peripheral run-off (at least two patent leg arteries) is also necessary. Nevertheless, further studies and longer follow-up are required to confirm the results obtained in our small series.