Limitations of in situ hybridization for the detection of bluetongue virus in blood mononuclear cells.

C. A. Dangler, A. de la Concha-Bermejillo, Bennie Osburn, Jeffrey L Stott

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

In situ nucleic acid hybridization was tested for the ability to detect bluetongue virus (BTV) nucleic acids in blood mononuclear cells. A standard protocol was devised and applied to the demonstration of BTV genetic sequences in cultured bovine mononuclear cells that had been infected in vitro. In situ hybridization using biotinylated single-stranded RNA probes, in the presence of 50% formamide at 50 C, demonstrated an intense, positive signal in 0.001-0.01% of the BTV-infected cultured mononuclear cells. The protocol was applied to isolated mononuclear cells from an experimentally infected heifer. No infected cells were observed by this method, although the blood specimens were obtained during peak viremia.

Original languageEnglish (US)
Pages (from-to)303-307
Number of pages5
JournalJournal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
Volume2
Issue number4
StatePublished - Oct 1990

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Bluetongue virus
in situ hybridization
In Situ Hybridization
Blood Cells
blood
Nucleic Acid Hybridization
RNA Probes
Viremia
cells
Nucleic Acids
Cultured Cells
RNA probes
viremia
nucleic acid hybridization
nucleic acids
heifers
cattle

ASJC Scopus subject areas

  • Microbiology
  • veterinary(all)

Cite this

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abstract = "In situ nucleic acid hybridization was tested for the ability to detect bluetongue virus (BTV) nucleic acids in blood mononuclear cells. A standard protocol was devised and applied to the demonstration of BTV genetic sequences in cultured bovine mononuclear cells that had been infected in vitro. In situ hybridization using biotinylated single-stranded RNA probes, in the presence of 50{\%} formamide at 50 C, demonstrated an intense, positive signal in 0.001-0.01{\%} of the BTV-infected cultured mononuclear cells. The protocol was applied to isolated mononuclear cells from an experimentally infected heifer. No infected cells were observed by this method, although the blood specimens were obtained during peak viremia.",
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T1 - Limitations of in situ hybridization for the detection of bluetongue virus in blood mononuclear cells.

AU - Dangler, C. A.

AU - de la Concha-Bermejillo, A.

AU - Osburn, Bennie

AU - Stott, Jeffrey L

PY - 1990/10

Y1 - 1990/10

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