Sensitive non-radioactive nucleic acid hybridization assay for plum pox virus detection

L. Palkovics, J. Burgyán, E. Balázs

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

A new non-radioactive sandwich hybridization assay was designed to simplify the analysis of a large number of plant samples. Plant material was homogenized in 0.5% SDS and added directly to the hybridization reaction, in which a pair of identifying probes were used. One of the probes was biotinylated capture RNA specific for plum pox virus (PPV) strain SK-68; the other RNA probe was synthesized from a plasmid bearing the adjacent sequence of this strain and was labelled with digoxigenin (DIG). Both purified viral RNA and crude extracts from PPV-infected plants were used as target for sandwich hybridization. The hybridization reaction was carried out in a streptavidin-coated ELISA plate. After extensive washing, the viral RNA was detected by conventional colour reaction using anti-DIG/alkaline phosphatase conjugate. In comparative experiments, we have shown that this non-radioactive detection system is more sensitive than conventional ELISA techniques and we were able to detect virusspecific RNA in more than 50% of the ELISA-negative samples.

Original languageEnglish
Pages (from-to)387-392
Number of pages6
JournalResearch in Virology
Volume145
Issue numberC
DOIs
Publication statusPublished - 1994

Fingerprint

Plum Pox Virus
Nucleic Acid Hybridization
Digoxigenin
Enzyme-Linked Immunosorbent Assay
Viral RNA
RNA
RNA Probes
Streptavidin
Complex Mixtures
Alkaline Phosphatase
Plasmids
Color

Keywords

  • Biotin, Digoxigenin, Streptavidin, Sensitivity, Potyvirus
  • ELISA, RNA-RNA hybridization, Plum pox virus

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Sensitive non-radioactive nucleic acid hybridization assay for plum pox virus detection. / Palkovics, L.; Burgyán, J.; Balázs, E.

In: Research in Virology, Vol. 145, No. C, 1994, p. 387-392.

Research output: Contribution to journalArticle

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