Non-invasive prenatal diagnosis of multiple endocrine neoplasia type 2A using COLD-PCR combined with HRM genotyping analysis from maternal serum.

The multiple endocrine neoplasia type 2A (MEN2A) is a monogenic disorder characterized by an autosomal dominant pattern of inheritance which is characterized by high risk of medullary thyroid carcinoma in all mutation carriers. Although this disorder is classified as a rare disease, the patients affected have a low life quality and a very expensive and continuous treatment. At present, MEN2A is diagnosed by gene sequencing after birth, thus trying to start an early treatment and by reduction of morbidity and mortality. We first evaluated the presence of MEN2A mutation (C634Y) in serum of 25 patients, previously diagnosed by sequencing in peripheral blood leucocytes, using HRM genotyping analysis. In a second step, we used a COLD-PCR approach followed by HRM genotyping analysis for non-invasive prenatal diagnosis of a pregnant woman carrying a fetus with a C634Y mutation. HRM analysis revealed differences in melting curve shapes that correlated with patients diagnosed for MEN2A by gene sequencing analysis with 100% accuracy. Moreover, the pregnant woman carrying the fetus with the C634Y mutation revealed a melting curve shape in agreement with the positive controls in the COLD-PCR study. The mutation was confirmed by sequencing of the COLD-PCR amplification product. In conclusion, we have established a HRM analysis in serum samples as a new primary diagnosis method suitable for the detection of C634Y mutations in MEN2A patients. Simultaneously, we have applied the increase of sensitivity of COLD-PCR assay approach combined with HRM analysis for the non-invasive prenatal diagnosis of C634Y fetal mutations using pregnant women serum.

Introduction of an automated system for the diagnosis and quantification of hepatitis B and hepatitis C viruses.

Hepatitis B virus (HBV) and Hepatitis C virus (HCV) infections pose major public health problems because of their prevalence worldwide. Consequently, screening for these infections is an important part of routine laboratory activity. Serological and molecular markers are key elements in diagnosis, prognosis and treatment monitoring for HBV and HCV infections. Today, automated chemiluminescence immunoassay (CLIA) analyzers are widely used for virological diagnosis, particularly in high-volume clinical laboratories. Molecular biology techniques are routinely used to detect and quantify viral genomes as well as to analyze their sequence; in order to determine their genotype and detect resistance to antiviral drugs. Real-time PCR, which provides high sensitivity and a broad dynamic range, has gradually replaced other signal and target amplification technologies for the quantification and detection of nucleic acid. The next-generation DNA sequencing techniques are still restricted to research laboratories.The serological and molecular marker methods available for HBV and HCV are discussed in this article, along with their utility and limitations for use in Chronic Hepatitis B (CHB) diagnosis and monitoring.

Application of maldi-tof mass spectrometry in clinical virology: a review.

MALDI-TOF mass spectrometry is a diagnostic tool of microbial identification and characterization based on the detection of the mass of molecules. In the majority of clinical laboratories, this technology is currently being used mainly for bacterial diagnosis, but several approaches in the field of virology have been investigated. The introduction of this technology in clinical virology will improve the diagnosis of infections produced by viruses but also the discovery of mutations and variants of these microorganisms as well as the detection of antiviral resistance. This review is focused on the main current applications of MALDI-TOF MS techniques in clinical virology showing the state of the art with respect to this exciting new technology.

Técnicas de imagen : proceso de soporte

Publicado en la página web de la Consejería de Salud: www.juntadeandalucia.es/salud (Consejería de Salud / Profesionales / Nuestro Compromiso por la Calidad / Procesos de Soporte)

Evaluación del cartílago articular mediante técnicas de dignóstico por la imagen

This article briefly reviews the composition of the hyaline cartilage and its ultrastructure. Subsequently, we offer a brief review of the role of imaging techniques in the assessment of this pathology. These include the most useful pulse sequences for the morphological assessment of cartilaginous injuries using MRI, as well as how these injuries appear in the images, at pre and post-surgical intervals. Lastly, we mention the developments in MRI that allow us to close in on the biochemical assessment of normal and pathological cartilage.

Artrodesis de tobillo: resultados comparativos de diferentes técnicas

INTRODUCTION: Ankle arthrodesis in an intervention that may be performed using a variety of surgical approaches. The joint may be stabilized using diferent systems, with or without bone graft. MATERIAL AND METHODS: Retrospective study that collected the results of 40 patients who underwent ankle arthrodesis; the complications that arose were analyzed, results according to the AOFAS clinical rating system, global patient satisfaction and the relationship with the articular approach route, the !xation system or the usage of grafts.RESULTS: 55% of the patients presented one or more complication that motivated a total of 12 re-interventions in 10 patients (excludes removal of osteosynthesis material). The absence of consolidation, which required a new arthrodesis was more frequent in patients where osteosynthesis was not used (two out of three cases) than in those using screws (four out of 24 cases), while there was no register of this in 13 patients in which intramedullary pin. The use of the distal extreme of the fibula or the iliac crest as bone graft has failed to improve the consolidation. Arthroscopic approaches avoid the appearance of soft tissue complications, although the consolidation disorders were more frequent. The average score on the AOFAS clinical rating system was 56.4±21.2 and 55% of the patients were satisfied. CONCLUSIONS: Although the case histories fail to provide significant results, it is a demanding intervention, with an elevated complication rate. Intramedullary pins seem to favor improved consolidation.