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United States Department of Agriculture

Agricultural Research Service

Research Project: MICROBIAL INTERACTIONS AND INTERVENTIONS TO REDUCE TRANSMISSION OF FOODBORNE PATHOGENS THROUGH POULTRY

Location: Food and Feed Safety Research

Title: Evaluation of repetitive extragenic palindromic-PCR and denatured gradient gel electrophoresis in identifying Salmonella serotypes isolated from processed turkeys

Authors
item Anderson, Phelue -
item Hume, Michael
item Byrd, James
item Hernandez, Charles
item Stevens, Scott -
item Stringfellow, Kendre -
item Caldwell, David -

Submitted to: Poultry Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: February 15, 2010
Publication Date: June 1, 2010
Repository URL: http://hdl.handle.net/10113/45797
Citation: Anderson, P., Hume, M.E., Byrd II, J.A., Hernandez Jr, C.A., Stevens, S., Stringfellow, K., Caldwell, D. 2010. Evaluation of repetitive extragenic palindromic-PCR and denatured gradient gel electrophoresis in identifying Salmonella serotypes isolated from processed turkeys. Poultry Science. 89:1293-1300.

Interpretive Summary: Salmonella has been reported as the leading foodborne pathogen in the US. A study was conducted to compare the usefulness of an automated molecular technique and a non-automated molecular technique as diagnostic tools for identifying types of Salmonella. Fifty-four Salmonella isolates from two turkey processing plants (A and B) were used for this comparison. There were more errors in the non-automated technique when compared to the automated technique. The results from the automated molecular technique were generated within an hour, while the non-automated technique required nearly a day to obtain results. The data suggested that the automated and non-automated molecular techniques are useful tools for identifying types of Salmonella. In addition, the automated technique is more rapid, may have higher discriminatory power, but may be less cost effective than the non-automated technique. However, more research may be needed to validate this argument. Both techniques displayed high sensitivity in discriminating among types of Salmonella, and either method could be considered as an alternative to more expensive and time-consuming, conventional antibody-based typing systems. This information is of interest to researchers and producers of animal food sources desiring more economical, high-throughput, and faster Salmonella typing methods.

Technical Abstract: Salmonella has been reported as the leading foodborne pathogen in the US. A study was conducted to compare the use of automated repetitive extragenic palindromic (REP-PCR) and denaturing gradient gel electrophoresis (DGGE) as diagnostic tools for identifying Salmonella serotypes. The interspersed conserved repetitive sequence of the bacterial genome and the 16-23S rDNA intergenic spacer region were amplified for REP-PCR and DGGE, respectively. Fifty-four Salmonella isolates from two turkey processing plants (A and B) were used for this comparison: Brandenburg, Derby, Hadar, and Typhimurium, with n = 6, 21, 12, and 15, respectively. The REP-PCR was fully automated, whereas DGGE was run on an acrylamide gel, and the image was captured digitally. Both dendrograms were created using the unweighted pair group method with arithmetic average. There were more variations in percentage similarity in DGGE when compared to REP-PCR. The banding patterns were more distinct and uniform in the REP-PCR group than with DGGE. The results from the REP-PCR were generated within an hour, while the DGGE required nearly a day to run. The data suggested that DGGE and REP-PCR are useful tools for identifying Salmonella serotypes. In addition, REP-PCR is more rapid, may have a higher discriminatory power, but may be less cost effective than DGGE. However, more research may be needed to validate this argument. Both DGGE and REP-PCR displayed high sensitivity in discriminating among Salmonella serotypes, and either method could be considered as an alternative to more expensive and time-consuming, conventional antibody-based serotyping systems.

Last Modified: 9/2/2014
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