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http://dx.doi.org/10.9721/KJFST.2016.48.4.335

Comparison of DNA isolation methods for detection of foodborne pathogens by real-time PCR from foods  

Koo, Eun-Jeong (Department of Food and Nutrition, Kookmin University)
Kim, Dongho (Bio-Medical Science Co., Ltd.)
Oh, Se-Wook (Department of Food and Nutrition, Kookmin University)
Publication Information
Korean Journal of Food Science and Technology / v.48, no.4, 2016 , pp. 335-340 More about this Journal
Abstract
This study was conducted to find out the most suitable DNA isolation methods for PCR detection of foodborne pathogens. Four DNA isolation methods including Universal Genomic DNA Extraction Kit (TaKaRa), PrepMan Ultra (Applied Biosystems), boiling method and alkaline lysis method (w/PEG) were tested and compared. The Universal Genomic DNA Extraction kit (TaKaRa) was considered as the more efficient isolation method for Escherichia coli O157:H7 and Staphylococcus aureus in lettuce, fish and beef. Meanwhile to detect the foodborne pathogens directly from foods without enrichment, the four different buffers such as double-distilled water, saline, glycine-saline, glycine-saline with Tween-20 and beef extract were also evaluated. As a result, saline was more suitable buffer for E. coli O157:H7. And double-distilled water was more suitable buffer than saline for S. aureus, respectively
Keywords
DNA isolation; real-time PCR; pathogens; detection;
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