A multiplex reverse transcriptase-PCR assay for the genotyping of avian infectious bronchitis viruses.
Journal
Avian diseases
Journal Volume
54
Journal Issue
1
Pages
104-108
Date Issued
2010
Author(s)
Abstract
Infectious bronchitis viruses (IBVs) in Taiwan have been divided into two genogroups, Taiwan group I (TW-I) and Taiwan group II (TW-II). Heterologous Mass-type strains are widely used as vaccines in the field. This work reports on a rapid and reliable multiplex reverse transcriptase-polymerase chain reaction (mRT-PCR) assay for the genotyping of IBVs. Multiplex primer sets were designed to amplify the region covering hypervariable regions 1 and 2 of the S1 glycoprotein gene. Several local strains and commercially available vaccines were used for evaluating the viral genotyping assay, and a number of field isolates were examined for clinical application. The results showed that all of the examined IBVs were accurately genotyped by identifying the corresponding bands on agarose gels (TW-I: 322 bp, TW-II: 161 bp, Mass type: 256 bp) after the mRT-PCR, in agreement with the viral genome sequence data. The mRT-PCR assay was able to detect viral RNA copies as low as 10(3), 10(50, and 10(3) for the TW-I, TW-II, and Mass-type strains, respectively. The mRT-PCR assay accurately detected and discriminated vaccine viruses from wildtype strains in the field. This assay may be beneficial for virus identification and differentiation in routine disease surveillance.
Other Subjects
virus RNA; article; Avian infectious bronchitis virus; genetics; genotype; methodology; reproducibility; reverse transcription polymerase chain reaction; sensitivity and specificity; Genotype; Infectious bronchitis virus; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction; RNA, Viral; Sensitivity and Specificity
Type
journal article