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United States Department of Agriculture

Agricultural Research Service

Title: Recovery of Genotypic Information from Non-Culturable Campylobacter Jejuni Isolates in Wang's Transport Medium

Authors
item Callicott, Kenneth
item Stern, Norman
item Hiett, Kelli

Submitted to: Campylobacter Helicobacter and Related Organisms International Workshop
Publication Type: Abstract Only
Publication Acceptance Date: June 12, 2003
Publication Date: August 16, 2003
Citation: Callicott, K., Stern, N.J., Hiett, K.L. 2003. Recovery of genotypic information from non-culturable Campylobacter jejuni isolates in Wang's transport medium [Abstract]. International Journal of Medical Microbiology. 293:99.

Technical Abstract: Isolates of Campylobacter jejuni shipped internationally often arrive in a non-culturable state. We describe a polymerase chain reaction-based methodology whereby phylogenetic information can be recovered from non-culturable C. jejuni stored in Wang's transport medium. The robustness of this methodology was initially tested using five strains of C. jejuni isolated from various sources associated with poultry production. These strains were stored in Wang's transport medium before being subjected to one of five treatments designed to render the stored cells uncultivable: prolonged incubation at room temperature, prolonged incubation at 42°C, multiple rounds of freezing and thawing, boiling, or contamination with Pseudomonas aeruginosa (ATCC 27853). This method resulted in high molecular weight DNA, and amplicons of about 1.5 kb were easily obtained. A 1722 nucleotide section of the flaA locus was inconsistently obtained from the set of isolates as a whole, but all DNA preparations from any single isolate were consistently positive or negative, suggesting that a lack of amplification was due not to the method but to the genotype of the isolate. These results indicate that this method may be useful for strain typing schemes based on PCR amplification of Campylobacter DNA, including flaA short variable region (flaA SVR), multi-locus sequence typing (MLST), and flaA PCR-restriction fragment length polymorphisms (flaA PCR-RFLP). By using this method, isolates unrecoverable from "Wang's" can still be used to provide phylogenetic information for epidemiological studies.

Last Modified: 10/24/2014
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