Use of oligonucleotide probes for selecting potential high-yielding influenza reassortants.
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.
Sahasrabudhe
A
A
Biomolecular Research Institute, Parkville, Victoria, Australia.
Ekkel
S
S
Hampson
A W
AW
McKimm-Breschkin
J L
JL
D005814:Genes, Viral; D006801:Humans; D009980:Influenza A virus / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000138:chemical synthesis; D012367:RNA, Viral / Q000302:isolation & purification; D012324:RNA-Dependent RNA Polymerase; D016865:Reassortant Viruses / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification*; D013045:Species Specificity; D014763:Viral Matrix Proteins / Q000235:genetics; D017361:Viral Nonstructural Proteins / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics; D015678:Viral Structural Proteins / Q000235:genetics
D015345:Oligonucleotide Probes; C057027:PB2 protein, Influenzavirus A; D012367:RNA, Viral; D014763:Viral Matrix Proteins; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C077346:influenza virus polymerase basic protein 1; D012324:RNA-Dependent RNA Polymerase