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Hopp, H.E.; Hain, L.; Bravo Almonacid, F.; Tozzini, A.C.; Orman, B.; Arese, A.I.; Ceriani, M.F.; Saladrigas, M.V.; Celnik, R.; del Vas, M.; Mentaberry, A.N. "Development and application of a nonradioactive nucleic acid hybridization system for simultaneous detection of four potato pathogens" (1991) Journal of Virological Methods. 31(1):11-29
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Abstract:

cDNA clones of potato virus X (PVXcp strain), potato virus Y (PVYo strain), potato leaf roll virus (PLRV) and potato spindle tuber viroid (PSTV) were used separately or combined for the detection of the corresponding RNAs in extracts of infected plants. A general method for the rapid preparation of RNA extracts without use of organic solvents (i.e. phenol) was developed for this purpose. Plant extracts from a range of field, artificially inoculated germplasm genotypes, micro-propagated and protoplast samples, as well as vector insect extracts, were dot-blotted onto nylon or nitrocellulose membranes, subjected to sandwich nucleic acid hybridization with non-labelled specific single-stranded DNA probes followed by a biotin-labelled second step hybridization probe. Each probe was virus-specific but not strain-specific. Healthy or non-related plant extracts developed very faint or no signals. Sensitivity was tested by slot-blot hybridization. Detection levels were between 1.5 to 6 pg of viral nucleic acids and between 20 to 50 times more sensitive than standard double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). The assay developed was tested with material that was prepared for processing in the field (combination of fresh sap with extraction solution) and tested under simple laboratory conditions for detection. It was also successfully employed for screening of germplasm for virus resistance, detection of pathogens in vector insects, plantlets grown in vitro and in more sophisticated quantitative determinations of viral replication in artificially inoculated plants and protoplasts. © 1991.

Registro:

Documento: Artículo
Título:Development and application of a nonradioactive nucleic acid hybridization system for simultaneous detection of four potato pathogens
Autor:Hopp, H.E.; Hain, L.; Bravo Almonacid, F.; Tozzini, A.C.; Orman, B.; Arese, A.I.; Ceriani, M.F.; Saladrigas, M.V.; Celnik, R.; del Vas, M.; Mentaberry, A.N.
Filiación:Institute de Biologia Molecular, CICV-INTA Castelar, Argentina
Cátedra de Genética, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
Instituto de Ingeniería Genética y Biologia Molecular (INGEBI)-CONICET, Buenos Aires, Argentina
BioAlmidar Industrias Quimicas Almidar SA., Buenos AiresArgentina
Cátedra de Genética, la Facultad de Agronomia, Universidad de Buenos Aires, Buenos Aires, Argentina
Palabras clave:complementary dna; article; dna probe; dna rna hybridization; nonhuman; plant virus; potato; priority journal; rna virus; virus detection; Cloning, Molecular; DNA Probes; Enzyme-Linked Immunosorbent Assay; Methods; Nucleic Acid Hybridization; Plant Diseases; Plant Viruses; Potatoes; RNA, Viral; Sensitivity and Specificity; Support, Non-U.S. Gov't; Hexapoda; Insecta; Perodicticus potto; Potato leafroll virus; Potato spindle tuber viroid; Potato virus Y; RNA viruses; Solanum tuberosum; Viroids
Año:1991
Volumen:31
Número:1
Página de inicio:11
Página de fin:29
DOI: http://dx.doi.org/10.1016/0166-0934(91)90141-L
Título revista:Journal of Virological Methods
Título revista abreviado:J. Virol. Methods
ISSN:01660934
CODEN:JVMED
CAS:DNA Probes; RNA, Viral
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01660934_v31_n1_p11_Hopp

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Citas:

---------- APA ----------
Hopp, H.E., Hain, L., Bravo Almonacid, F., Tozzini, A.C., Orman, B., Arese, A.I., Ceriani, M.F.,..., Mentaberry, A.N. (1991) . Development and application of a nonradioactive nucleic acid hybridization system for simultaneous detection of four potato pathogens. Journal of Virological Methods, 31(1), 11-29.
http://dx.doi.org/10.1016/0166-0934(91)90141-L
---------- CHICAGO ----------
Hopp, H.E., Hain, L., Bravo Almonacid, F., Tozzini, A.C., Orman, B., Arese, A.I., et al. "Development and application of a nonradioactive nucleic acid hybridization system for simultaneous detection of four potato pathogens" . Journal of Virological Methods 31, no. 1 (1991) : 11-29.
http://dx.doi.org/10.1016/0166-0934(91)90141-L
---------- MLA ----------
Hopp, H.E., Hain, L., Bravo Almonacid, F., Tozzini, A.C., Orman, B., Arese, A.I., et al. "Development and application of a nonradioactive nucleic acid hybridization system for simultaneous detection of four potato pathogens" . Journal of Virological Methods, vol. 31, no. 1, 1991, pp. 11-29.
http://dx.doi.org/10.1016/0166-0934(91)90141-L
---------- VANCOUVER ----------
Hopp, H.E., Hain, L., Bravo Almonacid, F., Tozzini, A.C., Orman, B., Arese, A.I., et al. Development and application of a nonradioactive nucleic acid hybridization system for simultaneous detection of four potato pathogens. J. Virol. Methods. 1991;31(1):11-29.
http://dx.doi.org/10.1016/0166-0934(91)90141-L