Comparison of slot blot nucleic acid hybridization, immunofluorescence, and virus isolation techniques to detect bluetongue virus in blood mononuclear cells from cattle with experimentally induced infection.

A. de la Concha-Bermejillo, C. E. Schore, C. A. Dangler, C. C. de Mattos, C. A. de Mattos, Bennie Osburn

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

A slot blot hybridization technique was applied for detection of bluetongue virus (BTV) in blood mononuclear cells (BMNC) obtained from cattle with experimentally induced infection. This technique lacked sensitivity to detect the viral nucleic acid directly in clinical specimens. When aliquots of mononuclear cells from these cattle were cultivated in vitro for 10 days to amplify virus titer, only 33.3% of the samples collected during viremia gave a positive signal in the slot blot hybridization format. By contrast, results for 34.3% of noncultured and 63.3% of cultured mononuclear cell samples collected during viremia were positive by immunofluorescence. The average number of infected cells, as detected by immunofluorescence in the noncultured mononuclear cell samples, was 1 to 5/300,000, and was usually > 10/300,000 in the cultured cell samples. Virus was isolated from all postinoculation blood samples obtained from 4 heifers that were seronegative at the time of inoculation, but was not isolated from any of the preinoculation samples, or from any of the postinoculation samples obtained from 2 heifers that were seropositive at the time of inoculation. When virus isolation was attempted from separated mononuclear cells in 2 heifers, 43.7% of the noncultured and 87.5% of the cultured samples had positive results.

Original languageEnglish (US)
Pages (from-to)2245-2250
Number of pages6
JournalAmerican Journal of Veterinary Research
Volume53
Issue number12
StatePublished - Dec 1 1992

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Bluetongue virus
Nucleic Acid Hybridization
isolation techniques
nucleic acid hybridization
fluorescent antibody technique
Fluorescent Antibody Technique
Blood Cells
Viremia
Viruses
viruses
Cultured Cells
cattle
blood
Infection
infection
cells
Viral Load
heifers
sampling
Nucleic Acids

ASJC Scopus subject areas

  • veterinary(all)

Cite this

Comparison of slot blot nucleic acid hybridization, immunofluorescence, and virus isolation techniques to detect bluetongue virus in blood mononuclear cells from cattle with experimentally induced infection. / de la Concha-Bermejillo, A.; Schore, C. E.; Dangler, C. A.; de Mattos, C. C.; de Mattos, C. A.; Osburn, Bennie.

In: American Journal of Veterinary Research, Vol. 53, No. 12, 01.12.1992, p. 2245-2250.

Research output: Contribution to journalArticle

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abstract = "A slot blot hybridization technique was applied for detection of bluetongue virus (BTV) in blood mononuclear cells (BMNC) obtained from cattle with experimentally induced infection. This technique lacked sensitivity to detect the viral nucleic acid directly in clinical specimens. When aliquots of mononuclear cells from these cattle were cultivated in vitro for 10 days to amplify virus titer, only 33.3{\%} of the samples collected during viremia gave a positive signal in the slot blot hybridization format. By contrast, results for 34.3{\%} of noncultured and 63.3{\%} of cultured mononuclear cell samples collected during viremia were positive by immunofluorescence. The average number of infected cells, as detected by immunofluorescence in the noncultured mononuclear cell samples, was 1 to 5/300,000, and was usually > 10/300,000 in the cultured cell samples. Virus was isolated from all postinoculation blood samples obtained from 4 heifers that were seronegative at the time of inoculation, but was not isolated from any of the preinoculation samples, or from any of the postinoculation samples obtained from 2 heifers that were seropositive at the time of inoculation. When virus isolation was attempted from separated mononuclear cells in 2 heifers, 43.7{\%} of the noncultured and 87.5{\%} of the cultured samples had positive results.",
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