Detection and genotyping of human papillomavirus by real-time PCR assay

BackgroundDiagnosis of human papillomavirus (HPV) disease remains a challenge due to several factors related to the cost, the workload of available commercial assays to detect and genotype HPV, and to the low prevalence of infected patients.ObjectiveOur study aimed to develop a real-time PCR, based on SPF10 primers, in order to combine HPV-DNA detection and genotype identification avoiding the negative samples.Study designValidation of SYBR-green based SPF10 real-time PCR on HPV-DNA plasmids followed by the investigation of the viral status in 92 samples from oropharyngeal (94%) cutaneous biopsies (3%) and anal smears (3%) which had previously been HPV-genotyped by LiPA hybridization. In-house HPV viral loads were performed to evaluate the SPF10 real-time PCR sensitivity.ResultsData showed that 100% of HPV plasmids, assessable by LiPA hybridization, were detected and genotyped appropriately after SPF10 real-time PCR assays. These results defined a range of melting temperature peaks for HPV positivity by real-time PCR. The efficient determination of the presence of HPV-DNA by SPF10 real-time PCR was validated for 98% of clinical samples compared to commercial method. Discordant results were due to a low HPV-DNA amount and to a supplementary HPV genotype identified. The SPF10 real-time PCR sensitivity was evaluated between 1 and 10 copies/10³ cells using in-house HPV (6, 11 and 16) viral load assays.ConclusionThe real-time PCR method was efficient in combining screening and genotyping of HPV-DNA. Cost and workload reduction by SPF10 real-time PCR approach may facilitate earlier diagnosis and clinical management of HPV infected patients.     

Rural Surgery in Niger: A Multicentric Study in 21 District Hospitals

The purpose of the study was to evaluate the qualitative aspect and global impact of surgery in a district hospital (DH) since the launching of the surgery at the district level. Surgical care was provided by general practitioners (GP) who received 12-month training in surgery, certified by a “Capacity of District Surgery” (CDS) diploma. It was a prospective study during 4 years from 2007 to 2010. Of the 34 DHs, only 21 were functional and included in this study. Most of the DHs had two or more CDS (n?=?15). The majority of the DHs had one nurse surgical aid (n?=?16) and one nurse anesthetist (n?=?17). The total number of surgical operations was 18,441 cases; emergency cases represented 51.8 % and elective surgery 48.2 %. Regarding emergency surgery, cesarean sections revealed the most common surgical procedure (37.21 %), followed by wound debridement (19.42 %). In elective surgery, hernia repair and hydrocelectomy were the most common surgical procedures (69.60 %), followed by gynecologic procedures in 12.74 % of the cases. The global complication rate was 4.34 %. The global mortality rate was 1.04 % (n?=?192), 102 deaths following cesarean section (2.87 %). No death was encountered in elective surgery. Nine hundred and fifty-five patients (5.17 %) were transferred to a higher-level facility of whom 598 patients (62.61 %) were admitted for fracture treatment. The concept of district surgery has proven to be an effective tool to counter skilled medical manpower shortage to perform emergency and elective basic surgery at the rural level and could be adopted by developing countries facing similar health challenges.     

Paraneoplastic rheumatic syndromes: report of eight cases and review of literature

Malignant neoplasms are associated with a wide variety of paraneoplastic rheumatological syndromes. The paraneoplastic nature should be based on specific criteria. We report a series of eight cases of paraneoplastic rheumatic syndromes revealing an underlying neoplasia. Our series consists of six men and two women, with a mean age of 46.1 (20-69?years). The first case is a hypertrophic osteoarthropathy of Pierre Marie that occurred in a 20-year-old man 1?month after treatment for his nasopharyngeal carcinoma; the paraclinical examinations showed lung and bone metastasis. The second case is that of a bilateral shoulder-hand syndrome revealing an invasive squamous cell carcinoma of the cervix in a 63-year-old woman. The third case involved a 69-year-old patient who had surgery 2?years ago for prostate adenocarcinoma and presented with polymyalgia rheumatica revealing bone metastasis. We also report two cases of leukemia in adults revealed by polyarthritis. The sixth observation is that of a paraneoplastic scleroderma that occurred concomitantly with prostate cancer. The seventh case of an acute arthritis showed a B lymphoma. The eighth case is that of a 52-year-old patient who presented with inflammatory arthralgias, and digital clubbing revealing a squamous cell carcinoma of the skin. Paraneoplastic rheumatism remains a rare event, but knowledge of it is essential for early diagnosis of underlying cancer.