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Development of a PCR assay for identification of Bordetella hinzii

Register, Karen B.
Avian diseases 2013 v.57 no.2 pp. 307
Bordetella avium, Bordetella hinzii, DNA, Geographical Locations, bacteria, bird diseases, disease diagnosis, disease outbreaks, genes, immunocompromised population, nucleotide sequences, pathogens, polymerase chain reaction, poults, respiratory tract diseases, ribotypes, sequence analysis, turkeys
Bordetella hinzii infects primarily poultry and immunocompromised humans. Although initially thought to be nonpathogenic in poultry, it was recently shown that some strains cause disease in turkey poults indistinguishable from the clinical presentation of turkey coryza caused by Bordetella avium. B. hinzii and B. avium are closely related and share many genetic and phenotypic traits. Distinguishing between these bacteria is difficult and there is no method for identification of B. hinzii suitable for use by diagnostic laboratories. The ompA gene was identified as a potential target for a B. hinzii-specific PCR based on DNA sequence comparisons of amplicons from several candidate genes obtained from a limited number of B. hinzii and B. avium isolates. A PCR assay based on the ompA gene was designed for further testing. Assay sensitivity is 100% based on analysis of 48 B. hinzii isolates from diverse geographic locations representing all known ribotypes. Evaluation of 71 isolates of B. avium and 20 other bacterial isolates from poultry, comprising gram-negative and gram-positive commensals and pathogens of 9 genera, demonstrated an assay specificity of 100%. Reproducibility is 100% using either purified genomic DNA or cell lysates. The ompA PCR is a rapid, reliable and accurate method for identification of B. hinzii. It provides a valuable new tool for veterinary diagnostic laboratories investigating poultry respiratory disease outbreaks and may also be of value in the diagnosis of human infection.