Development and application of specific polymerase chain reaction assay targeting the gyrB gene for rapid detection of Riemerella anatipestifer

• Published in: Poultry science 2012-10, Vol.91 (10), p.2450-2453
• Main Authors: Wang, X. P., Zhu, D. K., Wang, M. S., Cheng, A. C., Jia, R. Y., Chen, S., Chen, X. Y., Tang, T.
• Format: Article
• Language: English
• Subjects: Animals; Bird Diseases - diagnosis; Bird Diseases - microbiology; DNA Gyrase - genetics; DNA Gyrase - metabolism; DNA, Bacterial - genetics; Ducks; Flavobacteriaceae Infections - diagnosis; Flavobacteriaceae Infections - veterinary; Liver - virology; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - veterinary; Riemerella - classification; Riemerella - genetics
• ISSN: 0032-5791; 1525-3171
• DOI: 10.3382/ps.2012-02375

ABSTRACT A pair of PCR primers was designed and synthesized to amplify a gyrB gene sequence from Riemerella anatipestifer (RA). A fragment of 194 bp was detected in RA-positive isolates, whereas other isolates were negative, which confirmed the high specificity of the primers and PCR conditions. As little as 1.6 × 104 cfu/mL of cultural liquid was required by this method. We compared a 16S rRNA sequence-based PCR method and a Biolog bacterial identification system used in the detection and identification of suspicious isolates of RA in clinical tests. The results showed that the gyrB-based PCR was consistent with the results of the Biolog identification system and was more specific. By applying the gyrB-PCR to detect RA strains in 56 duck livers, a positive rate of 46% (26/56) was observed, whereas the positive rate of 85 throat swabs from clinically healthy ducks was 11%. Thus, this method could be used for the epidemiological investigation and preliminary isolate identification of RA.