Co-Authors:
Dovas, C.I., Plant Pathology Laboratory, Faculty of Agriculture, Aristotle University of Thessaloniki, 540 06 Thessaloniki, Greece
Hatziloukas, E., Plant Pathology Laboratory, Faculty of Agriculture, Aristotle University of Thessaloniki, 540 06 Thessaloniki, Greece
Salomon, R., Plant Pathology Laboratory, Faculty of Agriculture, Aristotle University of Thessaloniki, 540 06 Thessaloniki, Greece
Barg, E., Plant Pathology Laboratory, Faculty of Agriculture, Aristotle University of Thessaloniki, 540 06 Thessaloniki, Greece
Shiboleth, Y., Plant Pathology Laboratory, Faculty of Agriculture, Aristotle University of Thessaloniki, 540 06 Thessaloniki, Greece
Katis, N.I., Plant Pathology Laboratory, Faculty of Agriculture, Aristotle University of Thessaloniki, 540 06 Thessaloniki, Greece
Abstract:
Reverse transcriptase (RT)-polymerase chain reaction (PCR) and immunocapture (IC)-RT-PCR protocols were developed and optimized for the sensitive detection of Onion yellow dwarf virus, Leek yellow stripe virus and allexiviruses infecting Allium species. Polyvalence of the designed primers was successfully demonstrated, using samples of different plant species and geographic origins. Different sample preparation procedures were evaluated for their suitability to provide appropriate PCR templates. IC-PCR, RT-PCR with plant tissue extracts, and RT-PCR with total RNA, proved to be 10 2-10 4 times more sensitive than double-antibody sandwich-enzyme-linked immunosorbent assay (ELISA). Furthermore, a 'one step' IC-RT-PCR assay was developed using plant leaf extract as template source, which proved to be 102 times more sensitive than ELISA, and convenient for testing large numbers of leaf samples.