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Quantitative Detection of Salmonella typhimurium Contamination in Milk, Using Real-Time PCR  

JUNG SUNG JE (Institute of Life Sciences & Resources and Department of Food Science & Biotechnology, Kyung Hee University)
KIM HYUN-JOONG (Institute of Life Sciences & Resources and Department of Food Science & Biotechnology, Kyung Hee University)
KIM HAE-YEONG (Institute of Life Sciences & Resources and Department of Food Science & Biotechnology, Kyung Hee University)
Publication Information
Journal of Microbiology and Biotechnology / v.15, no.6, 2005 , pp. 1353-1358 More about this Journal
Abstract
A rapid and quantitative real-time PCR was developed to target the invasion A (invA) gene of Salmonella spp. We developed quantitative standard curves based on plasmids containing the invA gene. Based on these curves, we detected Salmonella spp. in artificially contaminated buffered peptone water (BPW) and milk samples. We were able to determine the invA gene copy number per ml of food samples, with the minimum detection limit of $4.1{\times}10^{3}$ copies/ml of BPW and $3.3{\times}10^{3}$ copies/ml of milk. When applied directly to detect and quantify Salmonella spp. in BPW and milk, the present real-time PCR assay was as sensitive as the plate count method; however, copy numbers were one to two logs higher than the colony-forming units obtained by the plate count methods. In the present work, the real-time PCR assay was shown to significantly reduce the total time necessary for the detection of Salmonella spp. in foods and to provide an important model for other foodborne pathogens.
Keywords
Copy number; invA gene; real-time PCR; Salmonella;
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