title: Development and application of specific polymerase chain reaction assay targeting the gyrB gene for rapid detection of Riemerella anatipestifer.
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 × 10(4) 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.
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Sichuan, People's Republic of China.
authors.forename: X P
authors.initials: XP
authors.lastname: Wang
authors.forename: D K
authors.initials: DK
authors.lastname: Zhu
authors.forename: M S
authors.initials: MS
authors.lastname: Wang
authors.forename: A C
authors.initials: AC
authors.lastname: Cheng
authors.forename: R Y
authors.initials: RY
authors.lastname: Jia
authors.forename: S
authors.initials: S
authors.lastname: Chen
authors.forename: X Y
authors.initials: XY
authors.lastname: Chen
authors.forename: T
authors.initials: T
authors.lastname: Tang
mesh_terms: D000818:Animals; D001715:Bird Diseases / Q000175:diagnosis / Q000382:microbiology; D027081:DNA Gyrase / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics; D004372:Ducks; D045826:Flavobacteriaceae Infections / Q000175:diagnosis / Q000662:veterinary*; D008099:Liver / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D060494:Riemerella / Q000145:classification* / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; D027081:DNA Gyrase