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Development of PCR Method for Rapid Detection of Allergic Materials in Foods

PCR을 이용한 식품 중 알레르기 유발물질 검출법 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) ;
  • Shin, Jun-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) ;
  • 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 : 2012.11.27
  • Accepted : 2013.03.12
  • Published : 2013.06.30

Abstract

The method for detection foods containing allergenic materials by PCR was developed in this study. To detect allergenic raw material from processed food, species specific primer which up to 200bp for PCR product were designed or selected from advanced research. As target materials, 14 items were selected (12 target materials for allergen in Korea, 2 target materials for allergen in foreign countries). The amplicon size for eggs, milk, buckwheat, peanuts, beans, wheat, mackerel, crab, shrimp, pork, peach, tomato, almond, and sesame were confirmed 281, 131, 138, 120, 118, 127, 211, 174, 231, 138, 174, 132, 103, and 220bp, respectively. And any non-specific bands were not detected among each others. Detection method for allergenic material developed in this study could be used to investigate inaccurate goods for allergen labeling or non-intentional contaminant during processed foods manufacturing. In addition, the system will be usefully to detection accurate allergenic raw materials of export for other countries.

본 연구에서는 분자생물학적 방법을 통한 알레르기 유발 원재료 확인을 위해 PCR방법의 최적 조건을 구축하였다. 가공식품에서 알레르기 원료성분 확인을 위하여 200bp 내외의 PCR 산물을 생성할 수 있는 종특이 프라이머를 설계하거나 선행연구사업 결과를 활용하였다. 대상 원료로는 국내 식품알레르기 표시대상인 난류, 우유, 메밀, 땅콩, 대두, 밀, 고등어, 게, 새우, 돼지고기, 복숭아 및 토마토와 제외국에서 알레르기 유발 성분으로 규정하고있는 아몬드, 참깨를 포함하여 총 14종을 대상으로 하였다. 특이 프라이머를 사용하여 PCR 한 결과 난류, 우유, 메밀, 땅콩, 대두, 밀, 고등어, 게, 새우, 돼지고기, 복숭아, 토마토, 아몬드 및 참깨로부터 각각 281, 131, 138, 120, 118, 127, 211, 174, 231, 138, 174, 132, 103 및 220bp의 특이 밴드를 확인하였으며 상호간의 비 특이적 밴드는 검출되지 않았다. 본 연구에서 확립한 알레르기 유발 원재료 검출법은 식품의 부정확한 표시나 가공식품의 제조과정 중 알레르기 유발물질의 비의도적 혼입 등으로부터 소비자를 보호하고 향후 수출 제품에 있어서 정확한 알레르기 유발원재료 표기에 활용이 가능할 것으로 판단된다.

Keywords

References

  1. King, N., Helm, R. and Stanley, S.: Allergenic characteristics of a modified peanut allergen, Mol. Nutr. Food Res., 49, 963-971 (2005). https://doi.org/10.1002/mnfr.200500073
  2. Sampson, H.A.: Update on food allergy, J. Allergy Clin. Immunol., 113, 805-819 (2004). https://doi.org/10.1016/j.jaci.2004.03.014
  3. Frew, A.J.: Food allergy and intolerances, Allergy, 52, 675 (1997). https://doi.org/10.1111/j.1398-9995.1997.tb01052.x
  4. 한국소비자원 소비자위해감시시스템(http://www.kca.go.kr)
  5. 식품의약품안전청, 2005. 식품의약품안전청고시 제2005-12호.
  6. 식품의약품안전청, 2008. 일본의 알레르기 유발물질 검사법 규정: 번역집, 63.
  7. Roland, E.P. and Elke, A.K.: Polymerase Chain Reaction Techniques for Food Allergy Detection, J. AOAC Int., 87, 139 (2004).
  8. Hebert, P.D.N., Alina, C., Shelley, L.B. and Jeremy, R.: Biological identifications through DNA Barcodes, Proc. R. Soc. Lond. B., 270, 313-321 (2003). https://doi.org/10.1098/rspb.2002.2218
  9. Michael, T.M., Michael, B., Gregory, T.R. and Alfried, P.V.: DNA-based species delineation in tropical beetles using mitochondrial and nuclear markers, Philos. Trans. R. Soc., 360, 1925-1933 (2005). https://doi.org/10.1098/rstb.2005.1724
  10. Tomoaki, M., Atsushi, A., Takuma, T., Sumitaka, Y., Yutaka, O. and Yoshida, M.: Identification of animal species using the partial sequences in the mitochondrial 16S rRNA Gene, Legal Med., 11, 449-450 (2009). https://doi.org/10.1016/j.legalmed.2009.02.002
  11. http://www.inspection.gc.ca
  12. http://www.mhlw.go.jp
  13. http://www.fda.gov
  14. http://www.emea.europa.eu/ema
  15. Samuel, B., Lehrer, W., Elliott, H. and Gerald, R.: Why are some proteins allergenic? Implications for biotechnology, Crit. Rev. Food Sci. Nut., 36, 553-564 (1996). https://doi.org/10.1080/10408399609527739
  16. Lee, J.H., Song, K.Y., Hyeon, J.Y., Hwang, I.G., Kwak, H.S., Han, J.A., Chung, Y.H. and Seo, K.H.: Comparison of Standard Culture Method and Real-time PCR Assay for Detection of Staphylococcus aureus in Processed and Unprocessed Foods, Korean J. Food Sci. Ani. Resour., 30, 410-418 (2010). https://doi.org/10.5851/kosfa.2010.30.3.410
  17. Lee, S.Y., Jang, K.I., Woo, G.J., Kwak, H.S. and Kim, K.Y.: Development of Protocol for the Effective Detection of Feline Calicivirus as Norovirus Surrogate in Oyster and Lettuce, Korean J. Food Sci. Techinol., 39, 71-76 (2007).
  18. Park, Y.C., Ahn, C.Y., Jin, S.O., Lim, J.Y., Kim, K.H., Lee, J.H., Cho, T.Y., Lee, H.J., Park, K.S. and Yoon, H.S.: Identification of Raw Materials in Processed Meat Products by PCR Using Species-Specific Primer, J. Fd Hyg. Safety, 27, 68-73 (2012). https://doi.org/10.13103/JFHS.2012.27.1.068
  19. Rakoczy-Trojanowska, M. and Bolibok, H.: Characteristics and comparison of three classes of microsatellite-based maskers and their application in plants, Cell Mol Biol Lett., 9, 221-238 (2004).
  20. Park, Y.C., Lim, J.Y., Kim, M.R., Park, Y.E., Lim, J.D., Hwang, C.R., Kim, K.H., Lee, J.H., Cho, T.Y., Lee, H.J., Lee, S.J. and Han, S.B.: Identification of Fauly Red Pepper Powder Containing Seasoned Red-pepper Sauce, J. Fd Hyg. Safety, 27, 182-187 (2012). https://doi.org/10.13103/JFHS.2012.27.2.182
  21. Wunsch, A. and Hormaza, J.I.: Molecular characterization of sweet cherry (Prunnus avium L.) genotypes using peach [Prunnus persica (L.) Batsch] SSR sequences, Heredity, 89, 56-63 (2002). https://doi.org/10.1038/sj.hdy.6800101
  22. Shirasawa, K., Asamizu, E., Fukuoka, H., Ohyama, A., Sato, S., Nakamura, Y., Tabata, S., Sasamoto, S., Wada, T., Kishida, Y., Tsuruoka, H., Fujishiro, T., Yamada, M. and Isobe, S.: An interspecific linkage map of SSR and intronic polymorphism markers in tomato, Theor Appl Genet., 121, 731-739 (2010). https://doi.org/10.1007/s00122-010-1344-3
  23. 식품의약품안전평가원, 2012, 가짜식품 판별 검사 지침서(I) (2011).
  24. 식품공전, 제 10 일반시험법, 8 식품표시관련시험법 (2011).
  25. He, G., Ronghua, M., Newman, M., Gao, G., Pittman, R.N. and Prakash, C.S.: Microsatellites as DNA markers in cultivated peanut (Arachis hypogaea L.), BMC Plant Biology, 3 (2003).
  26. Wei, W., Qi, X., Wang, L., Zhang, Y., Hua, W., Li, D., Haixia, L. and Zhang X.: Characterization of the sesame(Sesamum indicum L.) global transcriptome using Illumina paired-end sequencing and development of ETS-SSR markers, BMC Genomics, 12, 451 (2011). https://doi.org/10.1186/1471-2164-12-451

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