Application of the PCR technique for a rapid, specific and sensitive detection of Salmonella spp. in foods.
We report here the use of a PCR based assay modified by us for the detection of Salmonella spp. in foods, based on amplification of a 236 bp Salmonella specific hin/H2 region [Way et al. (1993) Applied and Environmental Microbiology 59, 1473-1479], using Ampli Taq Gold polymerase. Using this assay we were able to detect all the Salmonella serovars tested. The limit of detection was 1 fg of purified target DNA or N x 10(0) (1-3 cells) cfu ml(-1) of pure bacterial culture. This assay could detect N x 10(0) cfu Salmonella cells g(-1) of the food sample unambiguously in presence of endogenous microflora following 6 h enrichment, thus requires a duration of approximately 10 h for the full processing from DNA template preparation, PCR and visualization of DNA product on agarose gel. The main advantage of this PCR detection method is its sensitivity, and specificity. We also tried to adopt DNA template isolation method simply by boiling the bacterial cells thereby reducing the possibility of contamination, cutting the processing time and cost considerably. This can be an added advantage for the use of this system in simple lab setups.
Agarwal
A
A
Industrial Toxicology Research Centre, PO Box 80, MG Marg, Lucknow-226 001, India.
Makker
A
A
Goel
S K
SK
D004254:DNA Nucleotidyltransferases / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific; D005506:Food Contamination / Q000032:analysis; D005516:Food Microbiology; D016133:Polymerase Chain Reaction / Q000592:standards*; D012475:Salmonella / Q000235:genetics*; D012680:Sensitivity and Specificity; D013997:Time Factors
D004269:DNA, Bacterial; D004254:DNA Nucleotidyltransferases; C059997:Hin recombinase; D015252:Deoxyribonucleases, Type II Site-Specific; C024403:GGCC-specific type II deoxyribonucleases