Detection of flavivirus RNA in infected cells using photobiotin-labelled hybridization probes.

Abstract:

:Ten plasmids containing viral cDNA inserts of portions of the dengue virus type 2 (DEN-2) or Kunjin virus (KUN) genomes were biotinylated using photobiotin acetate and used as probes for the detection of flavivirus RNA in infected Vero cells. The viral cDNA inserts ranged in length from 0.19 to 2.7 kilobase pairs, and represented segments of the flavivirus genome coding for structural and nonstructural proteins. In spot hybridization assays (hybridization at 60 degrees C) with RNA extracted from cells infected with one of fourteen different flaviviruses or Semliki Forest virus, all DEN-2 and KUN probes hybridized specifically with RNA from cells infected with DEN-2 or KUN, respectively. At the reduced stringency of lower temperatures, specific hybridization to homologous viral RNA was still a feature of the probes, and only limited cross-hybridization to the RNA of some other flavivirus species was detected.

journal_name

J Virol Methods

authors

Khan AM,Wright PJ

doi

10.1016/0166-0934(87)90055-3

subject

Has Abstract

pub_date

1987-02-01 00:00:00

pages

121-30

issue

2

eissn

0166-0934

issn

1879-0984

pii

0166-0934(87)90055-3

journal_volume

15

pub_type

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