RESUMO
The use of instrumental systems based on the surface plasmon resonance (SPR) for rapid diagnosis of intact plant viruses (in particular, tobacco mosaic virus (TMV)) is considered. A new approach using detection of viral antigen and antibody (IgG) complexes formed during the preincubation step (instead of their consecutive application in classical approach) is discussed. A comparison between signal level registered from the mixture of virus and specific serum and that from the sample without virus (samples deposited onto the sensor surface treated with thiocyanate and protein A Staphylococcus aureus) allows unambiguous detection of viral particles in the material studied. The performance capabilities of the method are discussed and illustrated by quantitative detection of virus in the actual samples (cells homogenate) at high concentration.
Assuntos
Anticorpos Antivirais/metabolismo , Complexo Antígeno-Anticorpo/análise , Vírus de Plantas/isolamento & purificação , Ressonância de Plasmônio de Superfície/métodos , Vírus de Plantas/imunologia , Proteína Estafilocócica A , Tiocianatos , Vírus do Mosaico do Tabaco/imunologia , Vírus do Mosaico do Tabaco/isolamento & purificaçãoRESUMO
The possibility has been demonstrated for applying a surface plasmon resonance for detecting plant viruses in real samples. An optimal mode for antiviral immunoglobulin immobilization on sensor surfaces is described. Out of three proposed techniques for sensor surface treatment, namely, unmodified gold surface, gold surface treated with (a) thiocyanate and (b) thiocyanate and protein A (Staphylococcus aureus), the latter was chosen as most suited for retention of the formed native immunoglobulin layer.