N. Lukacs, DETECTION OF VIRUS-INFECTION IN PLANTS AND DIFFERENTIATION BETWEEN COEXISTING VIRUSES BY MONOCLONAL-ANTIBODIES TO DOUBLE-STRANDED-RNA, Journal of virological methods, 47(3), 1994, pp. 255-272
Monoclonal antibodies to double-stranded RNA (dsRNA) are described for
use as a universal diagnostic tool to detect infection in plants by R
NA viruses. Crude nucleic acid extracts from plants infected with one
of 25 different viruses were examined by sandwich-ELISA and immunoblot
ting. In comparison to the corresponding controls elevated dsRNA conce
ntrations were found in 21 infected samples by ELISA; virus-specific d
sRNA bands from 18 viruses were detected by immunoblotting. Using this
method the identification of infecting virus is potentially possible
on the basis of the electrophoretic banding pattern of the dsRNA, whic
h in turn depends on the number, molecular weight and/or thermodynamic
stability of the dsRNA species present in the extract. Immunoblot ana
lyses in combination with temperature-gradient gel electrophoresis wer
e used to demonstrate that the four individual genomic dsRNAs of the c
oexisting beet cryptic viruses BCV1 and BCV2 can be distinguished from
one another and from other dsRNAs present in the extracts. It is show
n that the thermal denaturation profiles and the T-m-values of the mai
n structural transitions of BCV genomic dsRNAs are essentially the sam
e in viruses from sugar beet as well as from wild Beta maritima. The r
eliability of dsRNA-immunoblotting for detecting virus infection in pl
ants is discussed. Its use is especially recommended for the detection
and characterization of cryptic viruses.