Journal of Fermentation and Bioengineering, Vol.77, No.2, 137-143, 1994
Possible Use of a Polymerase Chain-Reaction Method for Specific Detection of Salmonella in Beef
A rapid and sensitive polymerase chain reaction (PCR) method for the detection of Salmonella isolates with different serotypes is described. Based on the DNA sequence of a cloned 1.8 kb HindIII DNA fragment which could hybridize with all the Salmonella isolates tested but not with any of the non-Salmonella isolates including Enterobacteriaceae closely related to Salmonella, oligonucleotide fragments ranging from 18- to 26-mer were synthesized and tested for their possible use as the PCR primers. Results showed that three oligonucleotides, called TS11, TS4 and TSS could be used in pairs of TS11/TS4 and TS11/TS5 for the PCR detection of salmonellae with various serotypes. Under the conditions described, non-Salmonella isolates did not generate any false positive results. For primers TS11/TS4, the molecular weight of the PCR product amplified with Salmonella DNA was 1179 bp while for primers TS11/TS5, the molecular weight amplified was 375 bp. Primer TS5 could also be used as a checking probe to identify the PCR product amplified from primers TS11/TS4. Study of the detection sensitivity showed that DNA from N x 10(0) or N x 10(1) cells of Salmonella could be detected unambiguously either with primers TS11/TS5 or with primers TS11/TS4. When these PCR primers were used for the detection of salmonellae in beef, N x 10(0)-N x 10(1) cells per gram of beef could be detected and the endogenously contaminated microflora in the food sample did not interfere with the detection.
Keywords:ENTEROTOXIGENIC STAPHYLOCOCCUS-AUREUS;LISTERIA-MONOCYTOGENES;DNA FRAGMENT;GROUND-BEEF;FOODS;PROBES