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.     

Fibrous Component of the Temporomandibular Joint Disk

This article describes the spatial arrangement of collagen fibrils in the articular disk of the normal human temporomandibular joint (TMJ). Disks obtained at necropsy were examined by light microscopy and scanning electron microscopy. Observations by light microscopy showed dense cartilaginous tissue composed of collagen fiber bundles running in all directions. Observations by scanning electron microscopy revealed a network of collagen fibrils which were predominantly organized in bundles. In the surface layers an anteroposterior orientation was detectable. The authors concluded that the fibrous component of the TMJ articular disk is composed of collagen fibrils, organized in a three-dimensional network and able to resist compression forces due to loading.     

Electromyography Before and After Treatment of TMJ Dysfunction

The patient described in this article suffered from pain in the right side of her face for five years before she saw the authors. Earlier treatment with medication and neurosurgery had not been successful. The authors used electromyography to examine the disturbed muscle function and to investigate a possible cause for the pain in the muscles of mastication. The silent periods of the patient's muscles were measured before and after treatment, since it is known from the literature that the EMG silent periods may be elongated in patients with TMJ dysfunction or occlusal irregularities. The coordination of both heads of this patient's lateral pterygoid muscles were bilaterally investigated before and after treatment. Treatment consisted of splint therapy and selective grinding. Oral rehabilitation was also undertaken, beginning three months after the disappearance of the patient's symptoms.     

Bilateral Lobar Lung Transplantation — the First Two Cases in Belgium

In the last twenty years lung transplantation has become an established treatment for end-stage lung failure refractory to medical management. Over this time, better short and long-term results have been achieved due to improvements in organ procurement, perfusion and preservation strategies, newer immunosuppressant regimes and better post-transplant care. The limiting factor for the number of lung transplantation procedures performed is the shortage of available donor organs. This results in longer waiting times for listed patients, with a substantially increased risk of dying prior to transplantation, especially in the paediatric population.

Several surgical strategies have evolved to overcome the donor shortage, with lobar transplantation becoming a viable alternative. We describe our initial experience with two young patients with end-stage cystic fibrosis (CF) who required lung transplantation. Given their small size it was not possible to transplant an entire lung from an adult donor in each hemithorax. We describe lobar transplantation as a technique used to overcome this, in the first such operation in Belgium.