RESUMO
The development of a quantitative-competitive reverse transcription-PCR (RT-PCR) assay to quantify dengue virus (DEN) genome (vRNA) and its replicative intermediate RNA (vRI) is described. A highly conserved region located on the DEN capsid-premembrane genes was used to produce a competitor RNA molecule which contains an internal deletion of 70 nucleotides. The competitor provides a suitable internal control useful to quantify viral RNA from all four dengue virus (DEN 1-4) serotypes. The detection limit of the assay was found to be 100 copies per reaction. This is a rapid, simple, sensitive, inexpensive and easy method for quantitation of DEN RNA species.
Assuntos
Vírus da Dengue/isolamento & purificação , Dengue/virologia , Genoma Viral , RNA Viral/análise , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Animais , Linhagem Celular , Dengue/diagnóstico , Vírus da Dengue/classificação , Vírus da Dengue/genética , Humanos , RNA Viral/sangue , Sensibilidade e Especificidade , SorotipagemRESUMO
Los anticuerpos surgieron en los organismos en respuesta a las necesidades imperantes de neutralizar y destruir los embates de agentes externos nocivos para los mismos. Los anticuerpos son macromoléculas que por sus propiedades de especificidad y afinidad a sus antígenos, han sido utilizados para toda una gama de estudios en la medicina, su manipulación fuera de los sistemas vivientes ha permitido su aplicación en la terapéutica y el diagnóstico oportuno de varias enfermedades. El presente trabajo muestra una sinopsis de las propiedades bioquímicas de los anticuerpos y de las estrategias más recientes que han permitido la manipulación de estas moléculas, con la finalidad de mejorar su afinidad y avidez, así como en los métodos de producción para incrementar su potencial de aplicación en la investigación biológica y médica.
Antibodies appeared in the organisms in response to the needs of neutralizing and destroying the attacks of external agents injurious to themselves. Antibodies are macromolecules that because of their properties of specificity and affinity to their antigens, have been used in a great variety of studies in medicine. Moreover, their manipulation out of living systems has permitted their application in the treatment and opportune diagnosis of several diseases. The present work shows a synopsis of antibodies biochemical properties and the most recent strategies that have allowed the manipulation of these molecules in order to improve their affinity and avidity. This work will also present the methodological advances that can increase antibodies application potential in biology and in medical research.