Bluetongue virus detection: a safer reverse-transcriptase polymerase chain reaction for prediction of viremia in sheep

J Vet Diagn Invest. 1997 Apr;9(2):118-24. doi: 10.1177/104063879700900202.

Abstract

A reversible target capture viral RNA extraction procedure was combined with a reverse-transcriptase nested polymerase chain reaction (PCR) to develop a capture PCR assay providing a rapid and safe prediction method for circulating bluetongue virus in infected ruminants. This new assay was compared with virus isolation and a recently developed antigen-capture enzyme-linked immunosorbent assay (ELISA) for the detection of bluetongue virus. Eight Warhill crossbred sheep were inoculated subcutaneously with bluetongue virus serotype 10, and blood samples were taken sequentially over a period of 28 days. The capture PCR detected the peak of viremia, as determined by virus isolation and antigen-capture ELISA, from day 5 to day 14 after challenge. The results indicate that the rapid-capture bluetongue virus PCR provides a rapid indicator of samples in which virus can be isolated. In addition, this capture bluetongue virus PCR procedure does not require a lengthy phenol extraction or the use of the highly toxic methyl mercury hydroxide denaturant.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Bluetongue / diagnosis*
  • Bluetongue virus / isolation & purification*
  • DNA Primers
  • Disease Susceptibility
  • Electrophoresis, Agar Gel
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Polymerase Chain Reaction / methods*
  • RNA, Viral / blood
  • RNA, Viral / isolation & purification
  • Sheep
  • Viremia / diagnosis*

Substances

  • DNA Primers
  • RNA, Viral