Emulsion PCR to improve sensitivity of PCR-based E-coli O157:H7 ATCC 35150 detection

초록

Conventional PCR (cPCR) with 38 thermal cycles (cPCR (38cycles) ) failed to amplify 3 copies of Escherichia coli O157:H7 DNA at the presence of 3.28x10(6) copies of Salmonella Typhimurium DNA, but emulsion PCR (ePCR) with 20 thermal cycles and a subsequent conventional PCR with 38 thermal cycles (ePCR (20cycles) -sPCR (38cycles) ) 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 3.28x10(6) 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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