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J Gen Virol 69 (1988), 991-998; DOI 10.1099/0022-1317-69-5-991
© 1988 Society for General Microbiology
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Detection of Potato Virus X by One Incubation Europium Time-resolved Fluoroimmunoassay and ELISA

Riivo Sinijärv, Lilian Järvekülg, Elena Andreeva and Mart Saarma

Department of Molecular Genetics, Institute of Chemical Physics and Biophysics, Academy of Sciences of the Estonian S.S.R., 200026 Tallinn, Estonian S.S.R., U.S.S.R.

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 x 104-fold and leaf extracts diluted up to 2 x 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.

Keywords: FIA, time-resolved, ELISA, one step incubation, PVX

Received 12 October 1987; accepted 18 January 1988.




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Copyright © 1988 by the Society for General Microbiology.