PCR amplification of the fimA gene sequence of Salmonella typhimurium, a specific method for detection of Salmonella spp.
The goal of this study was to evaluate the suitability of the fimA gene amplification by PCR as a specific method for detection of Salmonella strains. Salmonella typhimurium and other pathogenic members of the family Enterobacteriaceae produce morphologically and antigenically related, thin, aggregative, type 1 fimbriae. A single gene, fimA, encodes the major fimbrial unit. In order to obtain higher specificity, we have selected a series of primers internal to the fimA gene sequence and have developed a PCR method for detecting Salmonella strains. A collection of 376 strains of Salmonella comprising over 80 serovars, isolated from animals and humans in Canada, have been used to evaluate this PCR method. Forty non-Salmonella strains were also tested by the same procedure. Cultures were screened by inoculating a single colony of bacteria directly into a PCR mixture containing a pair of primers specific for the fimA gene. The specific PCR product is an 85-bp fragment which was visualized by polyacrylamide gel electrophoresis and ethidium bromide staining. All Salmonella strains gave positive results by the PCR. Feed and milk samples contaminated by Salmonella strains were also detected by this procedure. The detection of all Salmonella strains tested and the failure to amplify the fragment from non-Salmonella strains confirm that the fimA gene contains sequences unique to Salmonella strains and demonstrate that this gene is a suitable PCR target for detection of Salmonella strains in food samples.
Cohen
H J
HJ
Laboratory Services Division, Food Production and Inspection Branch, Agriculture and Agri-Food Canada, Ottawa, Ontario, Canada.
Mechanda
S M
SM
Lin
W
W
D001426:Bacterial Proteins / Q000235:genetics*; D015139:Blotting, Southern; D036781:Fimbriae Proteins; D016133:Polymerase Chain Reaction; D012486:Salmonella typhimurium / Q000235:genetics* / Q000302:isolation & purification; D012680:Sensitivity and Specificity
D001426:Bacterial Proteins; C063055:fimbrillin; D036781:Fimbriae Proteins