Characterization of infectious bronchitis virus isolates by slot blot hybridization.
We used slot blot hybridization of the hypervariable regions of the S1 subunit of spike peplomer gene to identify and characterize infectious bronchitis virus (IBV) strains. Template DNA was created from six reference strain IBVs of different serotypes and immobilized on a nitrocellulose membrane. We synthesized digoxigenin-labeled probes from reference and unknown field viruses and hybridized them to template DNA. All reference strains could be distinguished and isolates identified by serotype if they were at least 95% identical to a reference strain. This slot blot hybridization procedure was specific and reproducible, and strain typing was consistent with the S1 sequencing of the IBV genome. This study thus provides a simple and rapid method for typing of IBV.
Mondal
Shankar P
SP
Population Health and Reproduction, 1114 Tupper Hall, University of California, Davis, Davis, CA 95616, USA.
Cardona
Carol J
CJ
D000818:Animals; D001483:Base Sequence; D002645:Chickens; D017931:DNA Primers; D015342:DNA Probes; D018076:DNA, Complementary / Q000032:analysis / Q000737:chemistry; D004076:Digoxigenin; D001351:Infectious bronchitis virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity; D016415:Sequence Alignment / Q000662:veterinary; D012689:Sequence Homology, Nucleic Acid; D013698:Templates, Genetic
D017931:DNA Primers; D015342:DNA Probes; D018076:DNA, Complementary; D012367:RNA, Viral; D004076:Digoxigenin