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KMID : 1007520150240041559
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Food Science and Biotechnology
2015 Volume.24 No. 4 p.1559 ~ p.1563
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Emulsion PCR to Improve Sensitivity of PCR-based E. coli O157:H7 ATCC 35150 Detection
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Chai Chang-Hoon
Oh Se-Wook
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Abstract
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Conventional PCR (cPCR) with 38 thermal cycles (cPCR38cycles) failed to amplify 3 copies of Escherichia coli O157:H7 DNA at the presence of 3.28×106 copies of Salmonella Typhimurium DNA, but emulsion PCR (ePCR) with 20 thermal cycles and a subsequent conventional PCR with 38 thermal cycles (ePCR20cycles-sPCR38cycles) amplified them. Partitioning individual E. coli O157:H7 DNA with emulsion droplet improved the specificity of PCR and let 3 copies of E. coli O157:H7 DNA to be amplified specifically at the presence of 3.28×106 copies of S. Typhimurium DNA. Application of ePCR improved the sensitivity of PCR-based pathogen detection and enabled the detection of 3 copies of E. coli O157:H7 DNA.
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KEYWORD
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polymerase chain reaction, microdroplets, emulsion PCR, pathogen detection, PCR sensitivity
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