The use of PCR-based methodologies to characterize salmonella serotypes of poultry origin

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Title: The use of PCR-based methodologies to characterize salmonella serotypes of poultry origin
Author: Anderson, Phelue Nigel
Abstract: Three studies were conducted to investigate the use of molecular techniques to identify Salmonella serotypes in poultry . In the first experiment , two polymerase chain reaction (PCR ) -based techniques : denaturing gradient gel electrophoresis (DGGE ) and polyacrylamide gel electrophoresis (PAGE ) were used to analyze Salmonella serotype isolates from two turkey processing plants (A and B ) . Genotypic patterns of each isolate were compared with those of known serotypes identified by traditional antibody precipitation methods . In Plant A , four different Salmonella serotypes were identified : Derby , Hadar , Montevideo , and Senftenberg . In plant B , ten serotypes were identified : Agona , Anatum , Brandenburg , Derby , Hadar , Meleagridis , Montevideo , Reading , Senftenberg , and Typhimurium . S . Derby was predominant in Plant A (83 % ) while S . Typhimurium was the most common serotype recovered in Plant B (39 % ) . Overall , DGGE was more sensitive than PAGE . Isolates of the same serotypes were all grouped together by DGGE , while PAGE failed to group all like serotypes . Next , DGGE and REP -PCR were used as genotyping tools for identifying Salmonella . Fifty -four Salmonella isolates from two turkey processing plants (A and B ) were evaluated . The isolates were comprised of the following serotypes : Brandenburg , Derby , Hadar , and Typhimurium (n = 6 , 21 , 12 , and 15 , respectively ) . Both methods were very sensitive and detected diverse fingerprint profiles among the isolates . The data suggested that REP -PCR and DGGE are useful tools for identifying Salmonella serotypes in research trials of this type . The final trial was carried out to track Salmonella serotypes throughout an integrated poultry operation using DGGE . Four flocks were sampled from grow -out through processing . The data showed that there was correlation between Salmonella serotypes found on processed carcasses and during grow -out . In addition , the isolates were compared against 15 known serotypes in our data base and only S . Hadar from the data base matched the unknown Salmonella isolates . Overall , these studies demonstrate that PCR -based methods could be considered as an alternative to conventional methods of antibody -based serotyping . Molecular methods were found to be reliable , sensitive , inexpensive , reproducible , and less labor intensive than conventional methods .
URI: http : / /hdl .handle .net /1969 .1 /ETD -TAMU -3045
Date: 2009-05-15

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The use of PCR-based methodologies to characterize salmonella serotypes of poultry origin. Available electronically from http : / /hdl .handle .net /1969 .1 /ETD -TAMU -3045 .

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