Rapid and Specific Detection of Enterotoxigenic Escherichia coli and Salmonella Strains by Multiplex PCR Systems
| 發布日期：2008-05-14 | 維護日期：2023-03-08
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Rapid and Specific Detection of Enterotoxigenic Escherichia coli and Salmonella Strains by Multiplex PCR Systems
Shu-Jen Wang*
Department of Food Science & Technology, Chia Nan University of Pharmacy and Science,
60, Sec. 1, Erren Rd., Rende Township, Tainan County 717, Taiwan (R.O.C.)
(Received: February 5, 2007; Accepted: July 27, 2007)
Escherichia coli and Salmonella are two of the most important food-borne bacterial pathogens. Classical identification for these strains is laborious, time-consuming, and may generate erroneous results. The purpose of this study was to develop a rapid and specific multiplex PCR (m-PCR) method to simultaneously detect heat labile enterotoxin gene of E. coli (LT ETEC) and oriC of Salmonella sp. Multiplex PCR using two pairs of primers produced specific amplicons of expected sizes of 163 bp and 425 bp from mixed populations of Salmonella sp. and LT ETEC bacterial strains, respectively. These primers were then used for the detection of food and feces with 101-102 cells/g of Salmonella and LT ETEC, followed by SCLB (selenite cystine-lactose broth, selenite cystine / lactose broth, 5/3, w/w) incubation. The presence of these two pathogens in food and feces was detectable. Finally, we used this method for the detection of 160 kinds of market-available foods and feces, and found that LT ETEC bacterial strains were detected in 2 samples (poultry and feces), and one sample (feces) by the BAM (Bacteriological Analytical Manu) method.
and are two of the most important food-borne bacterial pathogens. Classical identification for these strains is laborious, time-consuming, and may generate erroneous results. The purpose of this study was to develop a rapid and specific multiplex PCR (m-PCR) method to simultaneously detect heat labile enterotoxin gene of (LT ETEC) and o of sp. Multiplex PCR using two pairs of primers produced specific amplicons of expected sizes of 163 bp and 425 bp from mixed populations of sp. and LT ETEC bacterial strains, respectively. These primers were then used for the detection of food and feces with 10-10 cells/g of and LT ETEC, followed by SCLB (selenite cystine-lactose broth, selenite cystine / lactose broth, 5/3, w/w) incubation. The presence of these two pathogens in food and feces was detectable. Finally, we used this method for the detection of 160 kinds of market-available foods and feces, and found that LT ETEC bacterial strains were detected in 2 samples (poultry and feces), and one sample (feces) by the BAM (Bacteriological Analytical Manu) method.
Key words: multiplex PCR, heat labile enterotoxin E. coli; Salmonella