Polymerase chain reaction and sequencing for typing rhinovirus RNA.
Primers were designed and tested for their ability to distinguish rhinoviruses from enteroviruses. A primer set derived from the 5'-UTR/VP coding 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 enterovirus RNA. It is shown that rhinoviruses could be specifically typed by sequencing the amplicon derived from the 5'-UTR set. The sequences 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 17,69, and 70 also group by sequence with the equivalent region of HRV14 from the genetic database, while the others group with 2 and 89.
Mori
J
J
Virus Reference Division, Central Public Health Laboratory, London, United Kingdom.
Clewley
J P
JP
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002522:Chlorocebus aethiops; D017931:DNA Primers; D004279:DNA, Viral; D004770:Enterovirus / Q000145:classification / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics*; D012229:Rhinovirus / Q000145:classification* / Q000235:genetics; D012680:Sensitivity and Specificity; D017421:Sequence Analysis / Q000379:methods; D012689:Sequence Homology, Nucleic Acid; D014709:Vero Cells
D017931:DNA Primers; D004279:DNA, Viral; D012367:RNA, Viral