Prevalence and genotyping of HPV, by cervical brushing, in Irpinia area of Campania region

Cervical cancer is due to persistent genital infection with Human Papillomavirus (HPV).The purpose of this study was to evaluate prevalence of HPV in Irpinia (Campania region, Italy), distribution of different viral genotypes, correlating cytological results and virological investigations. In the period 2006-2011, were made 1080 cervical samples of women aged 18-65 years for HPV identification and genotyping. Detection of the virus was performed by Multiplex-PCR System (Seegene,Arrow) and typing with INNO-LiPA HPV Genotyping Extra test (Innogenetics). Out of the 1080 tested samples, 330 (30.6%) samples were positive for HPV DNA. The most frequently occurring High Risk (HR)-HPV genotype in single infections was HPV16 (16.6%), followed by HPV51 (10.7%), in multiple infections HPV16 (15.7%) and 31 (14.6%). The prevalence of infection, correlated with age of patients studied, is greater in the group aged 26-30 years (42.5%). HR-HPV were detected in different percent in patients with Pap test scores: 22.5% in normal Pap smear (20% HPV16), 14.5% ASCUS (47.6% HPV16), 24% LSIL (20% HPV16), 79.3% HSIL (72.7% HPV16; 9.1% HPV18 detected only in this type of cellular alteration). The high prevalence of HR-HPV in patients with ASCUS or normal Pap test, suggesting the real advantage of HPV screening test, more sensitive in selecting the actual population at risk. Based on the findings of our epidemiological study, HR-HPV screening and HPV genotyping test should be strongly advised also to the vaccinated population for the high incidence of genotypes which are not included in vaccines (67%)

Reactivation of intestinal CMV in a renal transplant patient after 10 years from the transplant

Introduction.We analyzed the clinical case of a 51 years old man, kidney transplanted on December 2002. On April 2011, he had acute rectal bleeding, renal chronic rejection (creatinine 2.9 mg/dl), Hgb 8.7 g/dl, positive anti-CMV antibodies (IgG). A colonoscopy showed diverticulosis of the rectum associated with deepithelialisation. The patient was treated with maintenance immunosuppressive post-transplant therapy. On June 2011, the colonoscopy showed a stenosing lesion of the sigmoid colon, and blood sampling and intestinal biopsy were performed to search Cytomegalovirus (CMV) DNA by PCR. Methods. The presence of CMV-DNA was sought by automatic extractor QIACUBE, using QIAamp DNA BLOOD Mini Kit (Qiagen) for whole blood and QIAamp DNA Mini Kit (Qiagen) for biopsy.The extracted DNA was then amplified by Real Time PCR using Q-CMV RealTime Complete Kit (Nanogen), on instrument Applied Biosystems 7300. Results. At disease onset the viral load in whole blood was 208000 Geq/ml, and biopsy was positive. Antiviral therapy with Ganciclovir led to the negativity of the viral load and remission of symptoms. Conclusions. The clinical case described presented a reactivation of CMV infection in the intestine after more than 10 years from kidney transplantation, while the highest incidence of CMV reactivation usually occurs during the first year. In our opinion, the reactivation can be traced to long-term immunosuppressive therapy (maintenance posttransplant therapy) in combination with a state of inflammation of the intestinal mucosa. In fact, patients with IBD treated with steroid drugs, in particular the group of refractory to therapy and thus have a recovery of the inflammatory process, are exposed to reactivation of CMV with intestinal localization

Suspected chromosomally integrated human herpes virus 6 in hematopoietic stem cell transplantation

Background and aims: We report a case of a 27-year-old male affected by acute myeloid leukaemia MLL-PTD positive. After autologous stem cell transplantation, he was monitored based on cytomegalovirus, Epstein-Barr virus and human herpes virus 6 (HHV-6) DNA quantification in blood. Relapse occurred one year after transplantation; then the patient underwent to allogenic bone marrow transplantation using genotypically HLA-identical donor (sister). HHV-6 DNAemia was positive and persistently elevated, either after autologous either after allogenic transplant suggesting the occurrence of HHV-6 chromosomally integration. The work aim is to prove the occurrence of chromosomally integrated-HHV-6 (ci-HHV-6). Materials and Methods: HHV-6 DNA extraction was performed by automated extractor and DNA was amplified-quantified by Real Time polymerase chain reaction. Species identification was performed by sequencing HHV6-U100 glycoprotein using automated sequencer and sequencing products were analysed using the Blast program. Results: After autologous transplantation HHV6-DNAemia was 5.4 log copies/mL setting to 3.9 log copies/mL for a long period post allogenic transplantation. The patient’s hair follicles were tested for HHV- 6 DNA having positive results. Sequences of both strains of HHV6 extracts from blood and hair follicles resulted species B. HHV6 viral load decreased significantly after Lymphocyte Infusion by ci-HHV6 negative donor (sister), having steady viral load during the following six months of monitoring. One year later, patient is in complete haematological remission. Conclusions: Detection of HHV-6 in hair follicles and HHV-6 DNAemia persistently elevated before allogenic transplant, confirm the occurrence of ci-HHV-6. The observed important decreasing viral load is potentially due to the successful engraftment of ci-HHV-6-negative donor marrow after allogeneic transplant

Prevalence of Cytomegalovirus infections in ulcerative rettocolitis patients, using PCR in whole blood and biopsies

Background.The relationship between CMV and inflammatory bowel disease (IBD) is not yet fully understood; the viral reactivation could be related to the prolonged steroid treatment of patients with ulcerative colitis (UC). Study Design. Our study was conducted on 25 patients with reactivation of UC, all were screened for CMV research. CMV DNA was carried out by automatic extraction in whole blood and intestinal biopsies and amplified by RealTime PCR. Objectives.We assessed the prevalence of CMV infection and clinical outcome, particularly in CMV positive patients treated with antiviral therapy. Results. CMV DNA was detected in 11/25 patients (44%), 7 positive in both blood and biopsies, 2 positive biopsies, 2 positive in blood, but low charge. 14 CMV DNA negative patients: 8 had remission with conventional therapy, 6 treated with IFX (of which 2 underwent colectomy). 11 CMV DNA positive patients: 7 had remission with conventional therapy, 3 treated with IFX+Ganciclovir, 1 with Ganciclovir, obtaining negative viral load and clinical remission. Conclusion. The literature data available on the association CMV - IBD are partly conflicting. The decision on treatment with antiviral therapy of CMV positive patients is based on course of disease. On the other hand, some patients had remission without antiviral therapy. We considered viral load and resistance to conventional therapy to admit patients into antiviral treatment, getting acute phase remission. There are many unanswered questions regarding management of CMV reactivation in UC patients, which need long-term follow-up studies and larger population