Partial sequence identification of grapevine-leafroll-associated virus-1 and development of a highly sensitive IC-RT-PCR detection method

J Virol Methods. 2000 Apr;86(1):101-6. doi: 10.1016/s0166-0934(00)00135-x.

Abstract

Using immunocapture reverse transcription PCR (IC-RT-PCR) a specific PCR product from GLRaV-1 infected vine samples was amplified with the help of degenerate primers deduced from the conserved HSP70 region of closteroviruses. 511 basepairs of the 5'end of GLRaV-1 HSP70 gene were identified. Within this region, putative GLRaV-1 specific primers were designed and an IC-RT-PCR detection procedure was developed which is about 125 times more sensitive than the established ELISA method. No PCR product was amplified in GLRaV-2,-3 and -4 infected plants which indicates the specificity of the primers. This procedure may serve as an alternative method for GLRaV-1 detection where the sensitivity of ELISA is insufficient.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Closterovirus / genetics*
  • Closterovirus / isolation & purification*
  • DNA Primers
  • DNA, Complementary
  • HSP70 Heat-Shock Proteins / genetics*
  • Molecular Sequence Data
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Rosales / virology*
  • Sensitivity and Specificity
  • Sequence Alignment
  • Sequence Analysis, DNA

Substances

  • DNA Primers
  • DNA, Complementary
  • HSP70 Heat-Shock Proteins