Serotype identification of avian infectious bronchitis virus by RT-PCR of the peplomer (S-1) gene.
The S-1 peplomer gene sequences of 31 strains of avian coronavirus infectious bronchitis virus (IBV) from North America, Europe, and Australia were compared to identify common and unique regions for possible diagnostic applications. S-1 sequences that were conserved among serotypes and sequences that were variable between serotypes were identified. Based on conserved S-1 gene sequences, "general" degenerate oligonucleotide primers were designed that amplified IBV genomic RNA by the reverse transcriptase polymerase chain reaction (RT-PCR) procedure regardless of serotype. Primers specific for IBV serotypes Massachusetts, Connecticut, Arkansas, JMK, Delaware (DE/072/92), and California (CA/633/85) were designed from regions of the S-1 gene exhibiting extensive sequence hypervariability. The ability to identify these six serotypes of IBV by RT-PCR was demonstrated by testing the serotype-specific primers on a panel of unknown samples that included 30 reference strains and field isolates previously characterized by virus neutralization (VN). The use of serotype-specific primers in RT-PCR provides a rapid and accurate means of identifying IBV.
Keeler
C L
CL
Delaware Agricultural Experiment Station, Department of Animal and Food Sciences, College of Agricultural Sciences, University of Delaware, Newark 19717-1303, USA.
Reed
K L
KL
Nix
W A
WA
Gelb
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017124:Conserved Sequence; D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000737:chemistry; D001351:Infectious bronchitis virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009500:Neutralization Tests / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012367:RNA, Viral / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D012703:Serotyping / Q000662:veterinary; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics*
D017931:DNA Primers; D004279:DNA, Viral; D012367:RNA, Viral; D014759:Viral Envelope Proteins