“Cycliplex PCR” confirmation of Fusobacterium necrophorum isolates from captive wallabies: A rapid and accurate approach

Isolation and identification of obligate anaerobic bacteria is labour intensive and time consuming. This has led to the increased application of molecular tools to circumvent part of this problem. We report here the development of a rapid, accurate and cost-effective method to isolate and identify Fusobacterium necrophorum species from South Australian wallaby populations using a supplemented medium (BHIRS) in conjunction with a “Cycliplex PCR” method which involves a stepwise-selective amplification of target PCR products. This report demonstrates the complementation of phenotypic characterization by PCR for accurate and fast identification of F. necrophorum isolates from wildlife origin.

► Cycliplex PCR that targeted on housekeeping genes identifies Fusobacterium necrophorum.
► Cycliplex PCR uses a set of primer array for accurate characterization of isolates.
► Indole production: a reliable selection marker for potential F. necrophorum isolates.
► We develop BHIRS media that improved Fusobacterium sp. growth and indole production.
► We combine genotypic and phenotypic method for F. necrophorum characterization.