Submitted to: American Society for Microbiology
Publication Type: Abstract only
Publication Acceptance Date: 2/15/2005
Publication Date: 6/5/2005
Citation: Callicott, K., Hiett, K.L., Stern, N.J., Reiersen, J., Berndtson, E., Fridricksdottir, V., Gunnarsson, E., Georgsson, F., Lowan, R. 2005. Refinement of flaa svr sequence typing for campylobacter jejuni [abstract]. American Society for Microbiology. p. D-209. Interpretive Summary:
Technical Abstract: Background: Campylobacter jejuni is a common cause of bacterial food poisoning in the industrialized world, and the primary source of infection is from poultry. In order to better understand the molecular epidemiology of this bacterium in poultry and humans, one needs a reliable, reprooducible typing system. Meinersmann et al. (1997) described a method to amplify and sequence the short variable region (SVR) of flaA; we present a refinement of their method that greatly reduces the prevalence of ambiguous sequences we have observed. Methods: The FlaA SVR was amplified by the reverse primer (FIA625RU) of Meinersmann et al. and one of two forward primers: the original forward primer described by Meinersmann et al. (FIA242FU) and a second which more reliably amplified flaA alone (FLA4F). PCR products of the first primer set were sequenced for 543 Campylobacter isolates and for 487 isolates when the product was amplified by the second primer set. Results: Of the products generated by the first primer set, 580 (88%) were ambiguous. When the products generated using the second primer set were sequenced, 18 (3.6%) were ambiguous. Conclusions: When typeing C. jejuni using flaA SVR, the degree to which is ambiguous using primer FLA4F, one can sequence flaB using a forward primer in the 3 end of flaA and subtract that sequence from the ambiguous one.