Evaluation of four template preparation methods for polymerase chain reaction-based detection of Salmonella in ground beef and chicken.
OBJECTIVE
To compare procedures for recovering template DNA from ground beef or chicken for polymerase chain reaction (PCR)-based detection of Salmonella.


RESULTS
The primer set of ST11 and ST15 was utilized to amplify a 429-bp product from Salmonella serotype Typhimurium. Boiling and three commercial kits were evaluated for extracting DNA from pure suspensions and artificially contaminated ground beef and chicken. The detection sensitivity of the PCR assay for pure cultures was independent of the template preparation method (P=0.946). Boiling and GeneReleaser failed to detect Salm. Typhimurium at 4 x 106 cfu g(-1) in ground chicken. PrepMan Ultra and the high pure PCR template preparation kit facilitated reliable and sensitive detection of Salm. Typhimurium in two types of food. The sensitivities were approx. 4 x 103 cfu g(-1). When spiked samples were enriched in peptone water for 6 h, an initial inoculum of 1 cfu g(-1) was detectable.


CONCLUSIONS
Four template DNA preparation methods differed in performance with respect to the type of samples tested.


CONCLUSIONS
Template DNA for the PCR detection of pathogenic bacteria, such as Salmonella in meat and poultry, could be effectively obtained using a simple rapid method such as the commercially available PrepMan Ultra kit.
Li
Y
Y
Department of Food Science, University of Missouri, Columbia, MO 65211, USA.
Mustapha
A
A
D000818:Animals; D002417:Cattle; D002645:Chickens / Q000382:microbiology*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D008461:Meat Products / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012486:Salmonella typhimurium / Q000235:genetics / Q000302:isolation & purification*; D013698:Templates, Genetic
D017931:DNA Primers; D004269:DNA, Bacterial