Detection of Peanuts in Commercially Processed Foods by an Enzyme-Linked Fluorescent Immunoassay

Enzyme-linked fluorescent immunoassay에 의한 가공식품 중 땅콩의 검출

  • Published : 2009.02.28

Abstract

In this study we analysed for peanuts in processed foods using an enzyme-linked fluorescent immunoassay (ELFA), and compared the results with labeled ingredients. Crude peanut protein (CPP) was immunized into rabbits to produce specific antibodies(Ab). A sandwich ELFA was established using anti-CPP Ab and Ab-horseradish peroxidase (HRP) conjugate. The cross-reactivities of the Ab toward CPP, peanuts, almonds, soybeans, and walnuts were 100, 9.8, $1.1{\times}10^{-2},\;4.4{\times}10^{-3}$, and 0%, respectively. The samples included 19 items consisting of biscuits, snacks, chocolates, and so on. The results from the sandwich ELFA showed that peanuts were contained in 7 of the processed food items, among which, 5 items were labeled as having peanuts present but 2 items were not. One of the 2 items that was peanut-detected but unlabeled was a biscuit labeled to contain almonds and assayed to contain $2.1{\times}10^{-3}%$ peanuts, which might have been due to the weak cross-reactivity of the Ab toward almonds. The other item was a snack labeled to contain soybeans and assayed to contain 0.098% peanuts, which might have been due to peanut cross-contamination during processing, since the crossreactivity of the Ab toward soybeans was very weak. These results suggest that ELFA is a good tool to detect peanuts in processed foods, and allergens in certain processed foods should be labeled correctly.

Enzyme-linked fluorescent immunoassay(ELFA)를 이용하여 가공식품 중 땅콩을 분석하고, 그 결과가 표시내용과 일치하는지를 조사하였다. 땅콩으로부터 분리한 조단백질(CPP)을 토끼에게 면역하여 생산한 특이항체 및 horseradish peroxidase-항체 복합물을 이용하여 sandwich ELFA를 확립하였다. 특이항체의 교차반응은, CPP, 땅콩, 아몬드, 콩, 호도에 대하여 각각 100, 9.8, $1.1{\times}10^{-2},\;4.4{\times}10^{-3}$, 0%로 나타났다. 시료로는 비스킷, 스낵, 초콜릿 등 19점을 사용하였다. 이들의 분석결과, 7점의 시료에서 땅콩이 검출 되었으며, 이 중 땅콩의 함유가 표시된 제품은 5점이었고 그렇지 않은 제품은 2점이었다. 이들 2점 중 하나는 아몬드를 함유하고 있다고 표시된 비스킷으로 분석 상 미량($2.1{\times}10^{-3}%$)의 땅콩이 검출되었는데, 이는 교차반응 때문으로 추측된다. 나머지 1점은 대두의 함유가 표시된 스낵으로 분석 상 $9.8{\times}10^{-2}%$의 땅콩이 검출되었다. 이는 대두의 매우 낮은 교차반응율을 감안하면 제조공정상 땅콩의 교차오염 때문으로 추측된다. 이와 같이 ELFA에 의하여 가공식품 중 땅콩의 검출이 가능하였고, 현재 유통 되는 일부 가공식품 중 알레르겐의 표시를 정확히 할 필요가 있다고 생각한다.

Keywords

References

  1. Shon DH. Food and allergy. Food Sci. Ind. 33: 1-8 (2000) https://doi.org/10.1111/j.1365-2621.1968.tb00872.x
  2. Werfel T. Food allergy. J. Dtsch. Dermatol. Ges. 6: 573-583 (2008) https://doi.org/10.1111/j.1610-0387.2008.06778.x
  3. Host A, Haiken S. A prospective study of cow milk allergy in Danish infants during the first three years of life. Clinical course in relation to clinical and immunological type of hypersensitivity reaction. Allergy 45: 587-596 (1990) https://doi.org/10.1111/j.1398-9995.1990.tb00944.x
  4. Sampson HA, McCaskill C. Food hypersensitivity in atopic dermatitis evaluation of 113 patients. J. Pediatr. 107: 669-675 (1985) https://doi.org/10.1016/S0022-3476(85)80390-5
  5. Burks AW. Peanut allergy. Lancet 371: 1538-1546 (2008) https://doi.org/10.1016/S0140-6736(08)60659-5
  6. Sampson HA, Mendelson L, Rosen JP. Fatal and near-fatal anaphylactic reactions to food in children and adolescents. Engl. J.Med. 327: 380-384 (1992) https://doi.org/10.1056/NEJM199208063270603
  7. Korea Food and Drug Administration. Labeling Standards for Foods etc. KFDA Notification No. 2008-31 (Amendment on Jun.17, 2008) Available from:http://www.kfda.go.kr/open_content/data/food_view.php. Accessed Dec. 1, 2008
  8. Stephan O, Vieths S. Development of a real-time PCR and a sandwich ELISA for detection of potentially allergic trace amounts of peanut(Arachis hypogaea) in processed foods. J. Agr. Food Chem.52: 3754-3760 (2004) https://doi.org/10.1021/jf035178u
  9. Pomes A, Helm RM, Bannon GA, Burks AW, Tsay A, Champman MD. Monitoring peanut allergen in food products by measuring Ara h1. J. Allergy Clin. Immunol. 12: 640-645 (2003)
  10. Poms RE, Klein CL, Anklam E. Polymerase chain reaction techniques for food allergen detection. Food Addit. Contam. 21: 1-31 (2004) https://doi.org/10.1080/02652030310001620423
  11. Kiening M, Niessner R, Drs E, Baumgartner S, Krska R, Bremer M, Tomkies V, Reece P, Danks C, Immer U, Weller MG. Sandwich immunoassays for the determination of peanut and hazelnuttraces in foods. J. Agr. Food Chem. 53: 3321-3327 (2005) https://doi.org/10.1021/jf048394r
  12. Yolken RH, Stopa PJ. Enzyme-Linked Fluorescence Assay: Ultrasensitive solid-phase assay for detection of human rotavirus. J. Clin. Microbiol. 10: 317-321 (1979)
  13. Zaitsu K, Ohkura Y. New fluorogenic substrates for horseradish peroxidase: Rapid and sensitive assays for hydrogen peroxide and the peroxidase. Anal. Biochem. 109: 109-113 (1980) https://doi.org/10.1016/0003-2697(80)90017-2
  14. Laemmli UK. Cleavage of structural proteins during the assemblyof the head of bacteriophage T4. Nature 227: 680-685 (1970) https://doi.org/10.1038/227680a0
  15. Kim HJ, Shon DH, An enzyme-linked immunosorbent assay for detection of cooked goat meat. Korean J. Food Sci. Technol. 32:538-543 (2000)
  16. Tresca JP, Ricoux R, Pontet M, Engler R. Comparative activity of peroxidase-antibody conjugates with periodate and glutaraldehyde coupling according to an enzyme immunoassay. Ann. Biol. Clin. 53:227-231 (1995)
  17. Koppelman SJ, Wensing M, Ertmann M, Knulst AC, Knol EF. Relevance of Ara h1, Ara h2, and Ara h3 in peanut-allergic patients, as determined by immunoglobulin E western blotting, basophilhistamine release and intracutaneous testing: Ara h2 is the most important peanut allergen. Clin. Exp. Allergy 34: 583-590 (2004) https://doi.org/10.1111/j.1365-2222.2004.1923.x
  18. de Leon MP, Glaspole IN, Drew AC, Rolland JM, O'Hehir RE, Suphioglu C. Immunological analysis of allergenic cross-reactivity between peanut and tree nuts. Clin. Exp. Allergy 33: 1273-1280 (2003) https://doi.org/10.1046/j.1365-2222.2003.01761.x