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Competitive polymerase chain reaction (cPCR) was used to develop a direct enumeration method of Listeria monocytogenes in pork meat. Pork meat was artificially inoculated with L. monocytogenes and DNA was extracted using guanidine thiocyanate-phenol-chloroform and subjected to PCR amplification. Sixteen primer sets for L. monocytogenes hlyA gene were tested for sensitive detection and the DG69/DG74 primer set was selected. The detection limit achieved with this primer set was as low as 860 colony-forming units (cfu) per 0.1 g of pork meat. When the samples were cultured at $30^{\circ}C$ for 16 hr in Brain Heart Infusion (BHI) medium, even a single bacterium could be detected with this primer set by PCR. For cPCR, the hlyA gene, which features a 148 bp-deletion, was cloned in the pGEM-4Z vector. A known amount of competitor DNA which has the same primer binding sites was co-amplified with L. monocytogenes total DNA from the artificially inoculated pork meat. The cell-number determined by cPCR was approximately equal to cfu from the Most Probable Number (MPN) method. The whole procedure took only 5 hr.

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이 논문을 인용한 문헌 (7)

  1. 2006. "" Food science and biotechnology, 15(2): 324~327 
  2. 2006. "" Food science and biotechnology, 15(6): 967~974 
  3. Moon, Ae-Rie ; Choi, Weon-Sang 2007. "Rapid Enumeration of Salmonella spp. in Contaminated Pork Meat Using Competitive PCR" 한국식품위생안전성학회지 = Journal of food hygiene and safety, 22(4): 248~256 
  4. 2007. "" Food science and biotechnology, 16(4): 515~519 
  5. 2007. "" Food science and biotechnology, 16(3): 337~342 
  6. 2007. "" Food science and biotechnology, 16(3): 355~360 
  7. 2008. "" Food science and biotechnology, 17(4): 761~765 


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