Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
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Detection and Differentiation of Intentional and Unintentional Mixture in Raw Meats Using Real-time PCR

Real-time PCR을 이용한 식육원료의 의도적, 비의도적 혼입 판별법 개발

  • Kim, Kyu-Heon (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Kim, Mi-Ra (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Park, Young-Eun (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Kim, Yong-Sang (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Lee, Ho-Yeon (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Park, Yong-Chjun (Novel Food Division, Ministry of Food and Drug Safety) ;
  • Kim, Sang Yub (Hazardous Substances Analysis, Busan Regional Korea Food and Drug Administration) ;
  • Choi, Jang Duck (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Jang, Young-Mi (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety)
  • 김규헌 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 김미라 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 박영은 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 김용상 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 이호연 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 박용춘 (식품의약품안전처 신소재식품과) ;
  • 김상엽 (부산지방식품의약품안전청 유해물질분석팀) ;
  • 최장덕 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 장영미 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀)
  • Received : 2014.07.04
  • Accepted : 2014.10.08
  • Published : 2014.12.31
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Abstract

In this study, the detection method was developed using real-time PCR to distinguish 4 species (bovine, porcine, horse, and chicken) of raw meats. The genes for distinction of species about meats targeted at 12S rRNA and 16S rRNA parts in mitochondrial DNA. Probes were designed to have a 5' FAM and a TAMRA at the 3' end. This study is to develop 4 species-specific primer and probes about raw materials and real-time PCR on 10 strains to observe the products of non-specific signal for similar species. As a result, any non-specific signal were not detected among each other. Real-time PCR method was developed for quantitation and identification of intentional and unintentional mixture in ground mixed meat (The difference of $C_T$ value between intentional mixture and 100% meat: $${\leq_-}$$ cycles, The difference of $C_T$ value between unintentional mixture and 100% meat: $${\geq_-}$$ cycles). The detection and differentiation of intentional and unintentional mixture in this study would be applied to food safety management for eradication of adulterated food distribution and protection of consumer's right.

본 연구에서는 식육원료 4종(소, 돼지, 말 및 닭)을 각각 판별하기 위하여 real-time PCR을 이용한 판별법을 개발하였다. 종 판별을 위한 유전자로는 미토콘드리아 유전자 중 12S rRNA와 16S rRNA 부분을 대상으로 하였으며, 특이 프로브의 Reporter dye는 FAM, Quencher dye는 TAMRA로 설계하였다. 개발된 프라이머 및 프로브 세트로 유사종에 대한 비 특이적 signal의 생성 유무를 관찰하기 위하여 총 10종을 대상으로 특이성을 확인한 결과, 비특이적 signal은 확인되지 않았다. 식육원료에 대하여 real-time PCR을 통한 식육 판별 시 식육 혼합 방법에 따른 $C^T$값의 유의적인 차이는 없었다. 의도적 혼합 및 비의도적 혼입을 판별하기 위한 정량법 개발에서는 의도적 혼합의 경우 100% 식육과의 $C^T$ 값 차이가 4 cycle 이내이고, 비의도적 혼입일 경우 100% 식육과의 $C^T$ 값 차이가 6 cycle 이상이었다. 따라서 본 연구를 통하여 개발한 식육 원료의 의도적, 비의도적 혼입 판별법은 불량식품 유통 근절 및 소비자 인권보호에 크게 기여할 것으로 기대된다.

Keywords

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