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1 March 2010 A Multiplex Reverse Transcriptase–PCR Assay for the Genotyping of Avian Infectious Bronchitis Viruses
Hui-Wen Chen, Ching-Ho Wang
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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 103, 105, and 103 for the TW-I, TW-II, and Mass-type strains, respectively. The mRT-PCR assay accurately detected and discriminated vaccine viruses from wild-type strains in the field. This assay may be beneficial for virus identification and differentiation in routine disease surveillance.

Hui-Wen Chen and Ching-Ho Wang "A Multiplex Reverse Transcriptase–PCR Assay for the Genotyping of Avian Infectious Bronchitis Viruses," Avian Diseases 54(1), 104-108, (1 March 2010). https://doi.org/10.1637/8954-060609-Reg.1
Received: 9 June 2009; Accepted: 1 October 2009; Published: 1 March 2010
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