Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
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Development of Species-Specific PCR to Determine the Animal Raw Material

종 특이 프라이머를 이용한 동물성 식품원료의 진위 판별법 개발

  • Kim, Kyu-Heon (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) ;
  • Kim, Yong-Sang (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) ;
  • Jung, Yoo Kyung (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Lee, Jae-Hwang (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Chang, Hye-Sook (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 Team, Busan Regional 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) ;
  • 이호연 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 김용상 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 김미라 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 정유경 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 이재황 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 장혜숙 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 박용춘 (식품의약품안전처 신소재식품과) ;
  • 김상엽 (부산지방식품의약품안전청 유해물질분석팀) ;
  • 최장덕 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 장영미 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀)
  • Received : 2014.06.28
  • Accepted : 2014.10.22
  • Published : 2014.12.31
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Abstract

In this study, the detection method was developed using molecular biological technique to distinguish authenticity of animal raw materials. The genes for distinction of species about animals targeted at Cytochrome c oxidase subunit I (COI), Cytochrome b (Cytb), and 16S ribosomal RNA (16S rRNA) genes in mitochondrial DNA. The species-specific primers were designed by that Polymerase Chain Reaction (PCR) product size was around 200 bp for applying to processed products. The target 24 raw materials were 2 species of domestic animals, 6 species of poultry, 2 species of freshwater fishes, 13 species of marine fishes and 1 species of crustaceans. The results of PCR for Rabbit, Fox, Pheasant, Domestic Pigeon, Rufous Turtle Dove, Quail, Tree Sparrow, Barn Swallow, Catfish, Mandarin Fish, Flying Fish, Mallotus villosus, Pacific Herring, Sand Lance, Japanese Anchovy, Small Yellow Croaker, Halibut, Jacopever, Skate Ray, Ray, File Fish, Sea Bass, Sea Urchin, and Lobster raw materials were confirmed 113 bp ~ 218 bp, respectively. Also, non-specific PCR products were not detected in compare species by species-specific primers. The method using primers developed in this study may be applied to distinguish an authenticity of food materials included animal raw materials for various processed products.

본 연구에서는 식품 중 동물성 사용원료의 진위 판별을 위하여 분자생물학적 기법을 이용한 시험법을 개발하였다. 동물성 식품원료의 종 판별을 위한 유전자로는 미토콘드리아 DNA에 존재하는 COI, Cytb, 및 16S rRNA 유전자를 대상으로 하였으며, 가공식품에 적용하기 위하여 PCR 산물의 크기는 200 bp 내외가 되도록 종 특이 프라이머를 설계하였다. 대상종으로는 가축류 2종, 가금류 6종, 민물어류 2종, 해양어류 13종 및 갑각류 1종, 총 24종을 선정하였으며 종 특이 프라이머를 이용하여 예상되는 PCR 산물의 생성 유무를 확인하였다. PCR을 수행한 결과 토끼, 여우, 꿩, 집비둘기, 멧비둘기, 메추리, 참새, 제비, 메기, 쏘가리, 날치, 열빙어, 청어, 까나리, 멸치, 참조기, 넙치, 조피볼락, 홍어, 가오리, 말쥐치, 농어, 성게 및 바닷가재에 대하여 각각 156, 204, 152, 160, 113, 163, 167, 152, 165, 121, 136, 151, 178, 178, 146, 188, 177, 166, 179, 218, 188, 185, 127 및 172 bp에서 PCR 증폭 산물을 확인하였다. 그리고 프라이머 별로 비교종에서는 비특이적 PCR 산물(non-specific PCR product)은 생성되지 않았다. 본 연구에서 개발된 유전자 분석법을 이용하여 동물성 식품원료가 사용된 식품 원료 및 가공식품의 진위 판별에 활용이 가능할 것이며, 불량식품 근절에 크게 기여할 것으로 기대된다.

Keywords

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