RT-PCR-ELOSA tests on pooled sample units for the detection of virus Y in potato tubers

A highly sensitive RT-PCR protocol able to detect potato virus Y (PVY) in p ooled sample units (tubers) was developed. PVY-specific primers selected in the coat protein gene were found to amplify a 359 bp fragment from diluted crude extract of infected tubers. For the detection of the amplification p roducts, a colorimetric detection procedure in microtiter plates was establ ished. The amplicons are hybridized between a covalently linked capture pro be and a specific biotinylated detection probe ELOSA tests. This detection method detects at least 50 pg of virus per reaction for the four cultivars tested. The RT-PCR-ELOSA assay was adapted to pooled units in order to incr ease the sample size while reducing the number of tests. (C) 2001 Elsevier Science B.V. All rights reserved.