Abstract
Summary A time-resolved fluorimmunoassay (TRFIA) with europium-labelled monoclonal antibodies (MAbs) was used to detect potato virus X (PVX) at a concentration of 5 pg/ml, in potato tuber extracts diluted up to 7 × 104-fold and leaf extracts diluted up to 2 × 107-fold. When mixed with potato leaf sap, PVX was detected at 100 pg/ml. TRFIA with simultaneous incubation of antigen and labelled antibody (one incubation) was two orders of magnitude more sensitive for PVX detection than the conventional double antibody sandwich ELISA. The fluorescence signal in TRFIA with separate incubations of antigen and labelled antibody (two incubations) with MAbs was linearly related to the virus concentration between 1.6 ng/ml and 1000 ng/ml; the method can therefore be used for the quantification of PVX. PVX detection in plant specimens by one incubation ELISA with MAbs was generally 10 times more sensitive than the standard two incubation procedure. Europium TRFIA with MAbs to potato viruses was found to be a very sensitive method for the detection of PVX, potato virus M, potato virus S, potato virus Y and potato leaf roll virus.
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