HUI-WEN CHEN2018-09-102018-09-102010http://www.scopus.com/inward/record.url?eid=2-s2.0-77952300180&partnerID=MN8TOARShttp://scholars.lib.ntu.edu.tw/handle/123456789/355678Infectious 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.[SDGs]SDG3[SDGs]SDG10[SDGs]SDG16virus 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 Specificityjournal article10.1637/8954-060609-Reg.12-s2.0-77952300180