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Comparative Studies of A/Leningrad/134/57 Wild-type and 47-times Passaged Cold-adapted Mutant Influenza Viruses: Oligonucleotide Mapping and RNA-RNA Hybridization Studies

¹ Influenza Branch, Center for Infectious Diseases, Centers for Disease Control, Atlanta, Georgia 30333, U.S.A.
² The D.I. Ivanovsky Institute of Virology, Moscow 123098, U.S.S.R.
³ Institute for Experimental Medicine, Leningrad, U.S.S.R.
and⁴ Moscow Institute for Viral Preparations, Moscow 109088, U.S.S.R.

Direct biochemical evidence for the existence of mutations in five of the RNA segments of the A/Leningrad/134/57 cold-adapted 47th passage mutant as compared with its wild-type progenitor has been obtained using two techniques. T₁ oligonucleotide mapping of total viral RNA as well as of individual RNA segments revealed changes in RNAs 4, 5 and 6. Analysis of S₁ nuclease-treated RNA-RNA hybrids on polyacrylamide gels revealed changes in at least one of the polymerase genes as well as in RNAs 4, 5, 6 and 7. These findings provide a direct demonstration for the existence of multiple mutations in the cold-adapted mutant vaccine strain.

Keywords: influenza A virus, mutations, vaccine

Received 3 January 1985; accepted 13 May 1985.