Comparison of methods for virus detection in Allium spp.

被引:42
作者
Dovas, CI [1 ]
Hatziloukas, E [1 ]
Salomon, R [1 ]
Barg, E [1 ]
Shiboleth, Y [1 ]
Katis, NI [1 ]
机构
[1] Aristotelian Univ Thessaloniki, Fac Agr, Plant Pathol Lab, GR-54006 Thessaloniki, Greece
来源
JOURNAL OF PHYTOPATHOLOGY-PHYTOPATHOLOGISCHE ZEITSCHRIFT | 2001年 / 149卷 / 11-12期
关键词
Allexivirus; immunocapture-reverse transcriptase (RT)-polymerase chain reaction (PCR); Leek yellow stripe virus; one-step RT-PCR; Onion yellow dwarf virus; Allium viruses;
D O I
10.1046/j.1439-0434.2001.00705.x
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
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.
引用
收藏
页码:731 / 737
页数:7
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