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 pooled 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 products, a colorimetric detection procedure in microtiter plates was established. The amplicons are hybridized between a covalently linked capture probe 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 increase the sample size while reducing the number of tests.
Chandelier
A
A
Faculté Universitaire des Sciences Agronomiques de Gembloux, Unité de Phytopathologie, Passage des Déportés, 2 B-5030 Gembloux, Belgium.
Dubois
N
N
Baelen
F
F
De Leener
F
F
Warnon
S
S
Remacle
J
J
Lepoivre
P
P
D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D010947:Plants, Toxic; D017800:Potyvirus / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D011198:Solanum tuberosum / Q000821:virology*; D014026:Nicotiana; D014158:Transcription, Genetic
D017931:DNA Primers; D004279:DNA, Viral