DEVELOPMENT OF REAL TIME POLYMERASE CHAIN REACTION FOR DETECTION OF Salmonella typhimurium AND Salmonella enteritidis IN FISH
Abstract: Previously designed
endpoint PCR has been adapted for use with real time PCR to detect the presence
of Salmonella typhimurium and Salmonella enteritidis in fish. Optimization of a standard curve in
the presence of herring sperm DNA as background matrix indicated that the real
time PCR highly efficient with the Pearson coefficient of determination (R2)
value = 0.99937 and slope (M) value = -3.44. An enrichment method (overnight
culture) significantly increased (p<0.05) the sensitivity of real time PCR.
Comparison of real time PCR and the conventional isolation method based on
biochemical tests has been conducted. In terms of their sensitivity, real time
PCR and the conventional methods are not significantly different in the level
of confidence 95%. Both real time PCR with enrichment method and conventional
biochemical method can detect the presence of Salmonella spp. in spiked sample.
However the direct extraction method was only detecting the presence of
Salmonella in higher concentration. While the sensitivity both conventional and
real time PCR are similar, the real time PCR has an advantage to detect the
pathogen qualitatively and quantitatively depending on processing method.
Keywords: real time PCR,
Salmonella contamination, fish
Author: Tuti Hartati Siregar,
Jennifer Elliman, Leigh Owens
Journal Code: jpperikanangg120022
