Comparison of Methods for Virus Detection in Allium spp

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 success...

Full description

Saved in:
Bibliographic Details
Published in:Journal of phytopathology 2001-12, Vol.149 (11), p.731-737
Main Authors: Dovas, C. I, Hatziloukas, E, Salomon, R, Barg, E, Shiboleth, Y, Katis, N. I
Format: Article
Language:English
Subjects:
Allexivirus
Allium
Allium viruses
Biological and medical sciences
Enzyme-linked immunosorbent assay
Fundamental and applied biological sciences. Psychology
Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control
immunocapture-reverse transcriptase (RT)-polymerase chain reaction (PCR)
leaf extracts
Leek yellow stripe virus
Microbiology
one-step RT-PCR
Onion yellow dwarf virus
Phytopathology. Animal pests. Plant and forest protection
Plant viruses and viroids
reverse transcriptase polymerase chain reaction
RNA-directed DNA polymerase
viruses
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:0931-1785
1439-0434
DOI:10.1046/j.1439-0434.2001.00705.x