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Simultaneous quantification of two plant viruses by double-label time-resolved immunofluorometric assay
M Saarma1, L Järvekülg, I Hemmilä
1Laboratory of Molecular Genetics, Estonian Academy of Sciences, U.S.S.R.
Abstract:
For simultaneous and sensitive detection of two antigens in one sample, monoclonal antibody (MAb) to potato virus M (PVM) was labelled with a lanthanide Eu3+ and MAb to potato virus X (PVX) with another lanthanide Sm3+. A mixture of the labelled MAbs was used as a tracer. After performing the immunoreactions, the fluorescence of the dissociated lanthanides was measured at different wave-lengths in a time-resolved fluorometer to quantificate the PVX and PVM amount in a sample. Double-label time-resolved fluoroimmunoassay (TR-FIA) detected 1 ng/ml of each virus and was therefore more sensitive for simultaneous detection of PVX and PVM than reported for a single virus detection with double antibody sandwich ELISA (DAS-ELISA).