Evaluation of TaqMan PCR assay for detecting Salmonella in raw meat and shrimp.
We evaluated the TaqMan Salmonella amplification/detection kit from PE Applied Biosystems, which uses a polymerase chain reaction (PCR) assay for rapid detection of Salmonella in food samples. This system uses the 5' nuclease activity of Taq DNA polymerase, which digests an internal fluorogenic probe to monitor the amplification of the target gene. The system's sensitivity and specificity were evaluated using 42 serotypes of 68 Salmonella strains isolated from fecal samples from patients in Tokyo, Japan, and 39 non-Salmonella strains in 22 genera. There were no false-negative or false-positive results. This PCR assay can detect 3 CFU per PCR tube of Salmonella in pure culture (120 CFU/ml of TSB culture). PCR signals were attenuated with artificially contaminated shrimp, but a similar detection limit was obtained. TaqMan's performance was tested with 100 meat and chicken samples purchased from stores in Tokyo. Overall, two of the DNA extraction protocols (the Chelex and EnviroAmp methods) worked equally well, with some exceptions. Of the 100 samples analyzed, 10 were positive for Salmonella with both conventional culture methods and the kit and 89 were negative with both. One sample was negative by the culture method but positive by the kit assay. These results indicate that TaqMan is a reliable and rapid method for Salmonella analysis in the food industry. With this system, food samples can be analyzed for Salmonella in less than 20 h.
Kimura
B
B
Department of Food Science and Technology, Tokyo University of Fisheries, Minato, Japan. Kimubo@tokyo-u-fish.ac.jp
Kawasaki
S
S
Fujii
T
T
Kusunoki
J
J
Itoh
T
T
Flood
S J
SJ
D000818:Animals; D003470:Culture Media; D004269:DNA, Bacterial / Q000302:isolation & purification; D033364:Decapoda / Q000382:microbiology*; D005069:Evaluation Studies as Topic; D005243:Feces / Q000382:microbiology; D005516:Food Microbiology; D006801:Humans; D008460:Meat / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012475:Salmonella / Q000254:growth & development / Q000302:isolation & purification*; D012480:Salmonella Infections / Q000382:microbiology; D012486:Salmonella typhimurium / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012703:Serotyping; D019914:Taq Polymerase
D003470:Culture Media; D004269:DNA, Bacterial; D019914:Taq Polymerase