Dot hybridization detection of plum pox virus using 32P-labeled RNA probes representing non-structural viral protein genes.
J Virol Methods
; 30(2): 161-71, 1990 Nov.
Article
en En
| MEDLINE
| ID: mdl-2148174
ABSTRACT
A cDNA library covering the complete genome of plum pox virus strain D (PPV D) has been obtained, and an endonuclease restriction map derived from it. This map was superposed on the PPV genomic organisation map, established for a nonaphid transmissible strain of PPV (Maiss et al., 1989). This allowed us to select seven probes, corresponding to different regions on the PPV genome. These probes were tested in a dot-blot hybridization assay for the detection of PPV. Probes of various lengths (0.25 to 1.5 kb) were tested and those measuring at least 0.8 kb (4 of the 7 probes selected) proved to be the most sensitive. The detection limit was of about 5 pg of purified virus per assay. Probes representing non-structural viral protein genes were equally sensitive in detecting both serotypes D and M of PPV. The previously described probe pBPPV1 (Varveri et al., 1988), covering the coat protein gene of strain D, was less sensitive, when compared to the above probes, in detecting heterologous strains of PPV. The polyvalence of probes transcribed from non-structural viral protein genes was confirmed by screening isolates of PPV, collected in infected orchards in several Mediterranean countries.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Virus de Plantas
/
Sondas ARN
/
Proteínas Estructurales Virales
/
Proteínas del Núcleo Viral
/
Cápside
/
Genes Virales
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
J Virol Methods
Año:
1990
Tipo del documento:
Article
País de afiliación:
Francia