July
1999
, Volume
83
, Number
7
Pages
639
-
643
Authors
A. G.
Gillaspie
,
Jr.
, and
S. E.
Mitchell
,
USDA, ARS, Plant Genetic Resources Conservation Unit, Griffin, GA 30223-1797
; and
G. W.
Stuart
and
R. F.
Bozarth
,
Department of Life Sciences, Indiana State University, Terre Haute 47809
Affiliations
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Accepted for publication 23 March 1999.
Abstract
ABSTRACT
A highly sensitive reverse transcription-polymerase chain reaction (RT-PCR) method was developed to detect cowpea mottle carmovirus (CPMoV) in newly acquired germ plasm of Vigna spp. It detected virus in tissues diluted up to 10-9. The preferred primers were designed from the RNA replicase cDNA sequence of CPMoV. These primers were able to detect CPMoV in plants infected with 10 different isolates of the virus. There were no cross-reactions with either bean mild mosaic or melon necrotic spot carmoviruses or any of the common cowpea viral pathogens tested. The RT-PCR method was up to 105 times more sensitive than direct antigen coating enzyme-linked immunosorbent assay (DAC-ELISA) in detecting CPMoV. The RT-PCR method gave no false positive reaction as is sometimes seen with ELISA.
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ArticleCopyright
The American Phytopathological Society, 1999