Quantitative detection of Escherichia coli O157:H7 in ground beef by immunomagnetic separation and competitive polymerase chain reaction
A protocol based on immunomagnetic separation (IMS) and competitive polymerase chain reaction (C-PCR) was developed for quantitative detection of Escherichia coli 0157:H7 in ground beef. Internal standard sequences were synthesized by PCR with hybrid primers composed in part of original primers for C-PCR. A single concentration of internal standards was used for C-PCR after calibration with known amounts of target DNA. After a 6 h enrichment in Tryptic Soy Broth (TSB), C-PCR enabled quantitative detection of 0.5 to 5.5 CFU per gram of ground beef with Shiga-like toxin 1 and 2 (SLT 1 and 2) primers respectively.
Robert E. Levin and J. Guan. "Quantitative detection of Escherichia coli O157:H7 in ground beef by immunomagnetic separation and competitive polymerase chain reaction" Food Biotechnology 16.3 (2002): 155-166.
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