POLYMERASE CHAIN-REACTION AND SEQUENCING FOR TYPING RHINOVIRUS RNA

Primers were designed and tested for their ability to distinguish rhin oviruses from enteroviruses. A primer set derived from the 5'-UTR/VP c oding region junction was able to amplify all the rhinovirus serotypes tested. Enteroviruses were either not amplified by these primer pairs or produced a band of larger size that could easily be discriminated from the rhinovirus-specific product. In contrast, primers embedded in the 5'-UTR region alone were able to amplify both rhinovirus and ente rovirus RNA. It is shown that rhinoviruses could be specifically typed by sequencing the amplicon derived from this 5'-UTR set. The sequence s of the 5'-UTR region of ten previously unsequenced rhinoviruses were derived. The sequences obtained cluster into two groups: 1B, 41, 15, 30, 63, 31, 56, and 44; and 17, 69, and 70. Amplicons from serotypes 1 7, 69, and 70 also group by sequence with the equivalent region of HRV 14 from the genetic database, while the others group with 2 and 89. (C ) 1994 Wiley-Liss, Inc.