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Development of Detection Method for Oilfish (Ruvettus pretiosus and Lepidocybirium flavobrunneum) as a Food Materials not Usable in Foods

식품원료로 사용금지 대상인 기름치 (기름갈치꼬치 및 흑갈치꼬치) 판별법 개발

  • Park, Yong-Chjun (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Kim, Mi-Ra (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Jung, Yong-Hyun (Food Standardization Division, Food & Drug Administration) ;
  • Shin, Joon-Ho (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Kim, Kyu-Heon (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Lee, Jae-Hwang (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Cho, Tae-Yong (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Lee, Hwa-Jung (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Lee, Sang-Jae (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration) ;
  • Han, Sang-Bae (Scientific Food Investigation Team, Food Safety Evaluation Department, National Institute of Food & Drug Safety Evaluation, Food & Drug Administration)
  • 박용춘 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 김미라 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 정용현 (식품의약품안전청 식품기준과) ;
  • 신준호 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 김규헌 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 이재황 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 조태용 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 이화정 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 이상재 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀) ;
  • 한상배 (식품의약품안전청 식품의약품안전평가원 식품위해평가부 식품감시과학팀)
  • Received : 2012.06.04
  • Accepted : 2013.02.25
  • Published : 2013.03.31

Abstract

Since 1 June 2012, it is prohibited to sell oilfish as a food material but there are still many illegal cases of selling oilfish as if it is tuna or grilled Patagonian toothfish. So it is absolutely crucial to construct the system to distinguish the real food material from oilfish. There are two sorts of oil fish called Ruvettus pretiosus and Lepidocybirium flavobrunneum involved in Percifomes order and Gempylidae class. 16S DNA gene region in mitochondria was selected to design the specific primers. For design species-specific primer, the theoretical experiment were performed for the sequences of R. pretiosus, L. flavobrunneum, Thunnus thynnus, Thunnus albacores, Makaira mitsukurii and Xiphias gladius, registered at the Gene bank from the National Centre for Biotechnology Information, using BioEdit 7.0.9.0. program. Through the analysis of the result from experiments, it was possible to design the 4 kinds of primers to distinguish R. pretiosus and L. flavobrunneum. As a comparison group, 3 kinds of tuna and 4 kinds of billfishes were selected and experimental verification was performed. As a result, for R. pretiosus and L. flavobrunneum, R.P-16S-006-F/R.P-16S-008-R and L.F-16S-004-F/L.F-16S-006-R primers were selected eventually and PCR condition was established. In addition, 178bp and 238bp of PCR products were confirmed from the established condition and non-specific band was not amplified among similar species. Therefore, the species-specific primers developed in this study would be very useful and used in various ways such as internet shopping mall and illegal distributions with fast and scientific results.

기름치를 참치회 또는 메로구이로 판매하는 사례가 있으며 국내에서는 2012년 6월1일부터 식품원료로 판매가 금지되어 이를 판별하는 시험법 마련이 필요하다. 기름치는 농어목(Perciformes) 갈치꼬치과(Gempylidae)에 속하는 기름갈치꼬치(R. pretiosus)와 흑갈치꼬치(L. flavobrunneum)가 있으며 이를 판별하기 위한 종 특이 프라이머를 개발하기 위하여 미토콘드리아에 존재하는 16S DNA 유전자부위를 선정하였다. 그리고 미국 국립보건원에서 운영하는 유전자 은행(www.ncbi.nlm.nih.gov)에 등록되어있는 기름갈치꼬치, 흑갈치꼬치. 참다랑어, 황다랑, 청새치 및 황새치의 염기서열을 대상으로 BioEdit ver. 7.0.9.0 프로그램을 사용하여 비교 및 분석을 실시하였다. 분석을 통하여 기름갈치꼬치 및 흑갈치꼬치를 판별할 수 있는 각각 4종의 프라이머를 설계하였다. 설계된 프라이머에 대하여 대조군으로 다랑어 3종(참다랑어, 황다랑어, 눈다랑어) 및 새치류 4종(청새치, 황새치, 녹새치, 돛새치)에 대한 실험적 평가를 실시하였다. 그 결과 기름갈치꼬치에 대하여는 R.P-16S-006-F/R.P-16S-008-R, 흑갈치꼬치는 L.F-16S-004-F/L.F-16S-006-R 프라이머를 최종 선정하였으며, PCR 조건을 확립하였다. 확립된 조건에서는 각각 178bp 및 238bp의 PCR 산물을 확인하였으며, 유사종간의 비특이적 밴드는 형성되지 않았다. 따라서 본 연구에서 개발된 기름치를 판별할 수 있는 종 특이 프라이머는 인터넷쇼핑몰 또는 시중에 불법적으로 유통 가능성이 있는 제품을 신속하고 과학적으로 판별할 수 있어 식품안전관리에 활용도가 매우 클 것으로 기대된다.

Keywords

Acknowledgement

Supported by : 식품의약품안전평가원

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