Development of quantitative real-time PCR for virus determination after purification procedure using monolithic cromatographic supports (CIM®)

Isolation and concentration of viruses from plant material is time consuming and demanding process that could be shorten and simplify by using some chromatographic techniques. Previous experiments have shown that CIM(R) monolithic support is a chromatographic material that can be used for purification, separation and concentration of plant viruses. We were able to concentrate virus particles from extremely diluted samples in which viruses were undetectable by ELISA or PCR. According to results, CIM(R) monolithic supports can be used for detection of viruses in irrigation water, from were they can be concentrated and subsequently detected by serological or other method. Project that we present here is a logical continuation of previous work. Two model plant viruses will be used. For detection and quantification of model viruses concentrated on CIM(R) monolithic chromatographic supports we will develop highly sensitive real-time PCR method. Conditions for separations and concentrations of model viruses on CIM(R) supports will be optimized. If needed, CIM supports will also be optimized. We will use the procedures for detection and quantification of viruses in rivers and lakes. Application of accurate molecular detection methods will significantly contribute to the knowledge about CIM(R) monolithic support properties and will help to describe a nature of virus-support interaction. Understanding of these interactions will enable application of CIM (R) supports on the other fields of virology.