Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Development of Duplex PCR Method for Simultaneous Detection of Rabbit (Oryctolagus cuniculus) and Cat (Felis catus) Meats

Duplex PCR을 이용한 토끼(Oryctolagus cuniculus)와 고양이(Felis catus) 육류의 동시 검출법 개발

  • Hong, Yeun (Department of Food Science & Biotechnology and Institute of Life Sciences & Resources, Kyung Hee University) ;
  • Kim, Mi-Ju (Department of Food Science & Biotechnology and Institute of Life Sciences & Resources, Kyung Hee University) ;
  • Yang, Seung-Min (Department of Food Science & Biotechnology and Institute of Life Sciences & Resources, Kyung Hee University) ;
  • Yoo, In-Suk (Department of Food Science & Biotechnology and Institute of Life Sciences & Resources, Kyung Hee University) ;
  • Kim, Hae-Yeong (Department of Food Science & Biotechnology and Institute of Life Sciences & Resources, Kyung Hee University)
  • Received : 2015.09.16
  • Accepted : 2015.10.07
  • Published : 2015.12.31
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Abstract

A duplex polymerase chain reaction (PCR) detection method was developed to authenticate the use of cat and rabbit in food and to prevent unlawful distribution of illegally butchered meat in both domestic and imported food market. Species-specific primers were designed targeting mitochondrial cytochrome b gene. The sizes of PCR products were 191 bp for cat and 101 bp for rabbit, which were relatively small for better application of the detection method on processed foods. Specificities of primers were verified using 21 animal species including cat and rabbit. Limit of detection was examined by serial dilution of the sample DNA and confirmed as 0.005 ng for rabbit and 0.0005 ng for cat using Bioanalyzer. The developed duplex PCR method showed specificity and sensitivity in the identification of two target species.

국내 유통 식품 수입 식품 중 토끼와 고양이 고기의 혼입 여부를 알아내고 불법 도축된 고양이 고기를 토끼 고기나 다른 고기로 속여 판매하는 것을 방지하기 위해 토끼와 고양이를 동시에 검출할 수 있는 polymerase chain reaction (PCR) 법을 개발하였다. 토끼와 고양이의 종 특이 프라이머는 미토콘드리아의 cytochrome b 유전자를 대상으로 하였고 개발된 프라이머를 가공식품에 활용하는 것을 고려하여 PCR 산물의 크기는 토끼 101 bp, 고양이 191 bp로 최소화 하였다. 프라이머의 특이성은 총 21종의 동물을 대상으로 검토하였다. 개발된 검출법의 검출 한계는 시료 DNA를 희석하여 PCR과 Bioanalyzer로 확인한 결과 토끼는 0.005 ng, 고양이는 0.0005 ng이었다.

Keywords

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