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Development of Sandwich ELISA for the Detection of Shrimp in Processed Foods

가공식품 중 새우의 검출을 위한 샌드위치 ELISA의 개발

  • Do, Jeong-Ryong (Functionality Evaluation Research Group, Korea Food Research Institute) ;
  • Back, Su-Yeon (Functionality Evaluation Research Group, Korea Food Research Institute) ;
  • Shon, Dong-Hwa (Functionality Evaluation Research Group, Korea Food Research Institute)
  • 도정룡 (한국식품연구원, 기능평가연구단) ;
  • 백수연 (한국식품연구원, 기능평가연구단) ;
  • 손동화 (한국식품연구원, 기능평가연구단)
  • Received : 2014.04.14
  • Accepted : 2014.07.25
  • Published : 2014.10.31

Abstract

A sandwich enzyme-linked immunosorbent assay method (sELISA) for detecting the presence of shrimp in processed foods was developed using rabbit polyclonal antibodies against tropomyosin produced by black tiger prawns (shrimp). Based on the standard curve derived using this method, the detection range of shrimp was determined to be $1-100{\mu}g/mL$. The cross-reactivity of these antibodies toward black tiger prawns, fleshy prawns, cocktail prawns, lobster, and blue crab was 100, 73, 155, 18, and 0%, respectively. When shrimp was heated for 10 min, the mean assay recovery of tropomyosin was 121-221% at $70-100^{\circ}C$ and 7.8% at $121^{\circ}C$. When shrimp was added to cream soup, weaning food, sausage, fish paste, and sauce, the mean assay recovery was 397, 639, 168, 234, and 0%, respectively. In sample tests involving 14 commercial items, the coincidence ratio of assay results and reference was 79%.

가공식품 중 새우의 검출을 위한 샌드위치 ELISA(sELISA)의 조건을 확립하기 위해 홍다리 새우의 tropomyosin에 대한 특이항체를 이용하여 하였다. 이때, 새우의 검출범위는 1-100 ppm (${\mu}g/mL$)이며, 검출한계는 0.3 ppm이었다. 특이항체의 교차반응 결과, 홍다리새우, 흰다리 새우, 칵테일 새우, 바다가재, 꽃게에 대한 반응성은 각각 100, 73, 155, 18, 0%를 나타내었으며 새우에 대한 특이성이 매우 높았다. 열처리한 시료와 항체간의 반응성은 $100^{\circ}C$까지는 121-221%로 안정하였으나 $121^{\circ}C$에서는 반응성이 급격히 감소하였다(7.8%). 크림스프, 이유식, 소시지, 어묵, 소스에 대한 spike test에서 새우의 분석회수율은 각각 397, 639, 168, 234, 0%로 소스를 제외하고 매우 높게 나타났다. sELISA에 의하여 14점의 시판시료 중 새우의 함유 유무를 조사한 결과, 정성적으로 원료의 표시사항과 일치하는 비율은 79%이었다.

Keywords

References

  1. Holgate ST, Polosa R. Treatment strategies for allergy and asthma. Nat. Rev. Immunol. 8: 218-230 (2008) https://doi.org/10.1038/nri2262
  2. Kamath SD, Abdel Rahman AM, Komoda T, Lopata AL. Impact of heat processing on the detection of the major shellfish allergen tropomyosin in crustaceans and molluscs using specific monoclonal antibodies. Food Chem. 141: 4031-4039 (2013) https://doi.org/10.1016/j.foodchem.2013.06.105
  3. Sampson HA. Updata on food allergy. J. Allergy Clin. Immunol. 113: 805-819 (2004) https://doi.org/10.1016/j.jaci.2004.03.014
  4. Korea Food and Drug Administration. Labeling Standards for Foods etc. KFDA Notification No. 2011-67 (2011)
  5. Jeong BJ, Park KH, Lee HH, Kim KE, Koe SW, Lee KY. Identification and characterization of shrimp allergens in korea. Korean J. Asthma. Allergy Clin. Immunol. 17: 278-285 (1997)
  6. Leung PS, Chu KH, Chow WK, Ansari A, Bandea CI, Kwan HS, Nagy SM, Gershwin ME. Cloning, expression, and primary structure of metapenaeusensis tropomyosin, the major heat-stable shrimp allergen. J. Allergy Clin. Immunol. 94: 882-890 (1994) https://doi.org/10.1016/0091-6749(94)90156-2
  7. Daul CB, Slattery M, Reese G, and Lehrer SB. Identification of the major brown shrimp (Penaeus aztecus) allergen as the muscle protein tropomyosin. Int. Arch. Allergy Immunol. 105: 49-55 (1994) https://doi.org/10.1159/000236802
  8. Daul CB, Morgan JE, Waring NP, McCants ML, Hughes J, Lehrer SB. 'Crustacea allergy' immunologic evaluation shrimpallergic individuals. J. Allergy Clin. Immunol. 80: 716-722 (1987) https://doi.org/10.1016/0091-6749(87)90293-4
  9. Shon DH, Kim HJ, Bae GW, Kim SM. An enzyme-linked immunosorbent assay for detection of milk proteins in food. Korean J. Food Sci. Technol. 32: 564-569 (2000)
  10. Kwak BY, Ko SH, Park CW, Son DY, Shon DH. Development of enzyme-linked immunosorbent assay for glyphosate-tolerant soybeans. Korean J. Food Sci. Technol. 35: 366-372 (2003)
  11. Asensio L, Gonzalez I, Garcia T, Martin R. Detemination of food authenticity by enzyme-linked immnosorbent assay (ELISA). Food Control 19: 1-8 (2008) https://doi.org/10.1016/j.foodcont.2007.02.010
  12. Fuller HR, Goodwin PR, Morris GE. An enzyme-linked immunosorbent assay (ELISA) for the major crustacean allergen, tropomyosin, in food. Food Agr. Immunol. 17: 43-52 (2006) https://doi.org/10.1080/09540100600572651
  13. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685 (1970) https://doi.org/10.1038/227680a0
  14. Kim HJ, Shon DH. An enzyme-linked immunosorbent assay for detection of cooked goat meat. Korean J. Food Sci. Technol. 32: 538-543 (2000)
  15. Herrmann JE, Hendry RM, Collins MF. Factors involved in enzyme-linked immunoassay of viruses and evaluation of the method for identification of enteroviruses. J. Clin. Microbiol. 10: 210-217 (1979)
  16. Kamath SD, Abdel Rahman AM, Komoda T, Lopata AL. Impact of heat processing on the detection of the major shellfish allergen tropomyosin in crustaceans and molluscs using specific monoclonal antibodies. Food Chem. 141: 4031-4039 (2013) https://doi.org/10.1016/j.foodchem.2013.06.105
  17. Faeste CK, Plassen C. Quantitative sandwich ELISA for the determination of fish in foods. J. Immunol. Method. 329: 45-55 (2008) https://doi.org/10.1016/j.jim.2007.09.007

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