Identifying bovine respiratory syncytial virus by reverse transcription- polymerase chain reaction and oligonucleotide hybridizations

R. D. Oberst, M. P. Hays, K. J. Hennessy, L. C. Stine, J. F. Evermann, C. L. Kelling

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

An assay to identify tissue culture cells infected with bovine respiratory syncytial virus (BRSV) that utilizes reverse transcription (RT), the polymerase chain reaction (PCR), and a synthetic oligonucleotide hybridization probe has been developed. The RT-PCR assay uses a BRSV-specific negative-sense oligonucleotide primer to synthesize cDNA from a BRSV fusion protein mRNA template and another BRSV-specific oligonucleotide primer (positive sense) upstream from the negative-sense primer for PCR amplification. In the presence of mRNA templates of BRSV isolates originating from locations throughout the United States, the BRSV RT-PCR assay resulted in amplified products (381 bp) that were specific to BRSV, as demonstrated in hybridizations with a positive-sense oligonucleotide probe complementary to internal sequences and in sequence comparisons with the F protein of BRSV 391-2. In analysis of the BRSV RT-PCR assay with prototype strains of human RSV subgroups A and B, amplification of a similar 381-bp RT-PCR product was not evident, and no RT-PCR product hybridized with the internal probe. We conclude that the specific ability to amplify DNA sequences of BRSV F protein mRNA by RT-PCR and then to demonstrate the presence of the amplified product with a BRSV-specific oligonucleotide probe will greatly add to the speed, sensitivity, and specificity of BRSV diagnostics.

Original languageEnglish (US)
Pages (from-to)1237-1240
Number of pages4
JournalJournal of clinical microbiology
Volume31
Issue number5
DOIs
StatePublished - 1993
Externally publishedYes

ASJC Scopus subject areas

  • Microbiology (medical)

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