Submitted to: Journal of Clinical Microbiology
Publication Type: Peer reviewed journal
Publication Acceptance Date: 10/27/2005
Publication Date: 3/1/2006
Citation: Siragusa, G.R., Danyluk, M.D., Hiett, K.L., Craven, S.E., Wise, M. 2006. Molecular subtyping of poultry - associated clostridium perfringens isolated by rep-pcr. Journal of Clinical Microbiology. 44:1065-1073. Interpretive Summary: Clostridium perfringens (Cp) is a harmful bacterium that contaminates our food supply as well as cause disease fatal to poultry. Cp can have a highly negative impact on efficient poultry production as well as cause food poisoning. In this research we have utilized a method of determining a characteritic signature for individual cultures of Cp isolated from the various sources of poultry production. This method is known as rep-PCR. The end result of rep-PCR is a barcode-like pattern that is unique for every individual strain or variety of Cp. These signature barcodes aid in determining which varieties of Cp are important to target for elimination from poultry versus those that are less of a concern for human food safety and food production. Rep-PCR also can be used to track the source of food borne disease. This proof-of-concept leads the way to using this technique for Cp analysis.
Technical Abstract: Subtyping of poultry production derived C. perfringens isolates was accomplished using rep-PCR with Dt primers (repDt-PCR). A total of 48 isolates of C. perfringens were obtained from the different stages of the broiler production chain: two separate breeder farms that both supplied broiler chicks to a single hatchery that in turn provided chicks to a single growout farm then to a single processing plant. Isolates were taken from fecal, egg shell, fluff, and carcass rinse samples as part of a previously reported temporally linked epidemiological survey. All 48 isolates were all typable (100% typability) by repDt-PCR. This subtyping method was rapid, highly reproducible and discriminatory: Simpson's discriminatory index calculated to be 0.96 for groupings of >95% correlation. A subset of isolates from this same library (n=45) were previously subtyped by automated ribotyping and compared to the rep-PCR technique. Rep-PCR is a technology that has been recently developed into a semi-automated process for use without agarose gels and offers a technically feasible means to subtype this group of bacteria from specific epidemiological or production environment settings.