Development of a Rapid Foodborne-pathogen-detection Method Involving Whole-genome Amplification
2016
Seong, J.Y., Kookmin University, Seoul, Republic of Korea | Ko, Y.J., R and D Management Team, SolGent Co., Ltd., Daejeon, Republic of Korea | Myeong, H.K., R and D Management Team, SolGent Co., Ltd., Daejeon, Republic of Korea | Oh, S.W., Kookmin University, Seoul, Republic of Korea
In this study, polyethylene glycol (PEG) was used to improve DNA amplification efficiency during whole genome amplification (WGA). Amplification efficiency was determined by adding PEG with different molecular weights to the WGA reaction. The greatest increase in amplification efficiency was obtained with PEG 4,000 used at 1.5% concentration. Foodborne pathogenic DNA was amplified by WGA and quantitatively analyzed by real-time polymerase chain reaction. DNA of Salmonella serotype Typhimurium, Listeria monocytogenes, and Vibrio parahaemolyticus was amplified 7,777.01, 9,981.22, and 1,239.03 fold, respectively, by WGA. On adding PEG in the WGA reaction (i.e., enhanced WGA [eWGA]), 18-40-fold more DNA amplification was achieved. Thus, these analyses showed that foodborne pathogens, which are usually present at very low concentration in foods, can be detected by real-time PCR and WGA.
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