Comparison of Methods for Virus Detection in Allium spp.
- 1 December 2001
- journal article
- Published by Wiley in Journal of Phytopathology
- Vol. 149 (11-12) , 731-737
- https://doi.org/10.1046/j.1439-0434.2001.00705.x
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 stripevirus 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 102–104 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.Keywords
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