Use of oligonucleotide probes for selecting potential high-yielding influenza reassortants

J Virol Methods. 1997 Nov;68(2):139-45. doi: 10.1016/s0166-0934(97)00119-5.

Abstract

A new method for rapid screening of high-yielding reassortants of influenza virus, as candidates for vaccine production, is described. Oligonucleotide probes specific for all the parent genes of A/PR/8/34 (PR8), except the HA and the NA were designed based on database information available for different influenza strains. Digoxigenin labelled probes were tested by slot-blot hybridizations to purified RNA from a panel of A/PR/8/34 wild type and A/PR/8/34 reassortant viruses. The results show that the vast majority of reassortants selected for their high growth yield had acquired the non-structural (NS), matrix (M) and RNA polymerase 2 (PB2) genes from the PR8 parent. It is proposed that probes for these genes provide the potential for a simple and rapid procedure for selection of candidate high-yield reassortants for vaccine production.

MeSH terms

  • Genes, Viral
  • Humans
  • Influenza A virus / genetics*
  • Influenza A virus / growth & development
  • Influenza A virus / isolation & purification*
  • Nucleic Acid Hybridization
  • Oligonucleotide Probes* / chemical synthesis
  • RNA, Viral / isolation & purification
  • RNA-Dependent RNA Polymerase
  • Reassortant Viruses / genetics*
  • Reassortant Viruses / growth & development
  • Reassortant Viruses / isolation & purification*
  • Species Specificity
  • Viral Matrix Proteins / genetics
  • Viral Nonstructural Proteins / genetics
  • Viral Proteins / genetics
  • Viral Structural Proteins / genetics

Substances

  • Oligonucleotide Probes
  • PB2 protein, Influenzavirus A
  • RNA, Viral
  • Viral Matrix Proteins
  • Viral Nonstructural Proteins
  • Viral Proteins
  • Viral Structural Proteins
  • influenza virus polymerase basic protein 1
  • RNA-Dependent RNA Polymerase