[Polymerase chain reaction (PCR) method for the detection of Salmonella by using Chelex resin after enrichment culture].
Salmonella has been implicated as a important pathogen associated with acute gastroenteritis or food borne diseases in humans. Rapid and accurate diagnostic tests for Salmonella are needed by both the food industry and clinical laboratories. A pair of oligonucleotide primers from the phoE gene was used to develop a polymerase chain reaction (PCR) procedure to detect Salmonella spp. When Chelex-extracted DNA from the SBG enrichment culture were used as templates for the PCR amplification (E-Chx), the positive results were obtained with a few cells of Salmonella. This highly specific and sensitive technique gave an efficiency of 1.2 times (73 samples were positive) when tested against conventional culture method (62 samples were positive) using 125 fecal samples stored at 4 degrees C for 1-16 months. Based on the results obtained in this study, it appears that the enrichment-PCR method (E-Chx) using Chelex-extracted DNA described here can be useful to screen a large number of samples such as foods or environmental specimens for contamination of Salmonella.
Kobayashi
K
K
Laboratory of Microbiology, Osaka Prefectural Institute of Public Health.
Taguchi
M
M
Seto
K
K
D001483:Base Sequence; D005809:Genes, Regulator; D007475:Ion Exchange Resins; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D011137:Polystyrenes; D011145:Polyvinyls; D018272:Porins / Q000235:genetics*; D012475:Salmonella / Q000302:isolation & purification*
D007475:Ion Exchange Resins; D015345:Oligonucleotide Probes; D011137:Polystyrenes; D011145:Polyvinyls; D018272:Porins; C006960:chelex