Abstract
Fruit allergies have become more common in recent years, and are now a serious health problem. In this study, a multiplex PCR assay was used to detect potential fruit allergens causing food allergy labeling in Korea. For the detection of these allergens, specific primer pairs were designed to amplify the allergen-coding genes Cyclophilin (tomato), Mdtl 1 (apple), Pru p 2.01A (peach) and Pectin methylesterase inhibitor (kiwi), and primer pair targeting the 18S ribosomal RNA gene was additionally used as an endogenous control. Primer specificity was assessed with 23 plant species. A mixture of DNA from the four fruits was serially diluted and used to determine the sensitivity of the multiplex PCR assay, which was approximately 0.08 ng. Eleven commercial fruit products were evaluated to verify the applicability of the multiplex PCR assay. This assay is expected to be a specific and efficient method for detecting fruit allergens in foods.
Similar content being viewed by others
References
Cheng F, Wu J, Zhang J, Pan A, Quan S, Zhang D, Kim HY, Li X, Zhou S, Yang L. Development and inter-laboratory transfer of a decaplex polymerase chain reaction assay combined with capillary electrophoresis for the simultaneous detection of ten food allergens. Food Chem. 199: 799–808 (2016)
Choi YJ, Yoon JM, Chang YS, Oh JW. Sensitization to component antigens in acute idiopathic urticarial in children. Allergy Asthma Respir. Dis. 5: 331–335 (2017)
Christopher M, Webber MD, Ronald W, England MD. Oral allergy syndrome: a clinical, diagnostic, and therapeutic challenge. Ann. Allergy Asthma Immunol. 104: 101–108 (2010)
Fernandez-Rivas M, Benito C, Gonzalez-Mancebo E, de Durana MD. Allergies to fruits and vegetables. Pediatr. Allergy Immunol. 19: 675–681 (2008)
Garcia BE, Lizaso MT. Cross-reactivity syndromes in food allergy. J. Investig. Allergol. Clin. Immunol. 21: 162–170 (2011)
Garino C, De Paolis A, Coisson JD, Bianchi DM, Decastelli L, Arlorio M. Sensitive and specific detection of pine nut (Pinus spp.) by real-time PCR in complex food products. Food Chem 194:980–985 (2016)
Graziano S, Gulli M, Marmiroli N. Detection of allergen coding sequences of kiwi, peach, and apple in processed food by qPCR. J. Sci. Food Agric. 98: 3129–3139 (2018)
Han J, Wu Y, Huang W, Wang B, Sun C, Ge Y, Chen Y. PCR and DHPLC methods used to detect juice ingredient from 7 fruits. Food Control 25: 696–703 (2012)
Hubalkova Z, Rencova E. One-step multiplex PCR method for the determination of pecan and Brazil nut allergens in food products. J. Sci. Food Agric. 91: 2407–2411 (2011)
Kim MJ, Kim HY. Species identification of commercial jerky products in food and feed using direct pentaplex PCR assay. Food Control 79: 1–6 (2017)
Kim HJ, Ryu JO, Song JY, Kim HY. Multiplex polymerase chain reaction of identification of shigellae and four Shigella species using novel genetic markers screened by comparative genomics. Foodborne Pathog. Dis. 14: 400–406 (2017)
Kondo Y, Urisu A. Oral allergy syndrome. Allergol. Int. 58: 485–491 (2009)
Lee EJ, Ryu HK. A study on food allergy occurrence and parental perceptions of food allergen labeling system in school food service. Korean J. Community Living Sci. 27: 521–531 (2016)
Lee SC, Son YW, Sim DW, Park KH, Lee JH, Park, JW. Oral allergy syndrome associated with weed pollinosis. Allergy Asthma Respir. Dis. 4: 458–461 (2016)
Lee S. Food allergy in children: focus on IgE-mediated food allergy. J. Korean Med. Assoc. 60(3):242–248 (2017)
Linacero R, Ballesteros I, Sanchiz A, Prieto N, Iniesto E, Martinez Y, Pedrosa MM, Muzquiz M, Cabanillas B, Rovira M, Burbano C, Cuadrado C. Detection by real time PCR of walnut allergen coding sequences in processed foods. Food Chem. 202: 334–340 (2016)
Mafra I, Ferreira IM, Oliveira MBPP. Food authentication by PCR-based methods. Eur. Food Res. Technol. 277: 649–665 (2008)
Marchisotto MJ, Harada L, Blumenstock J, Bilaver L, Waserman S, Sicherer S, Boloh Y, Regent L, Said M, Schnadt S, Allen K, Muraro A, Taylor S, Gupta RS. Global perceptions of food allergy thresholds in 16 countries. Allergy 71: 1081–1085 (2016)
Pafundo S, Gulli M, Marmiroli N. Multiplex real-time PCR using SYBR GreenER™ for the detection of DNA allergens in food. Anal. Bioanal. Chem. 396: 1831–1839 (2010)
Pafundo S, Gulli M, Marmiroli N. Comparison of DNA extraction methods and development of duplex PCR and real-time PCR to detect tomato, carrot, and celery in food. J. Agric. Food Chem. 59: 10414–40424 (2011)
Park YC, Kim MR, Shin JH, Kim KH, Lee JH, Cho TY, Lee HJ, Lee SJ, Han SB. Development of PCR method for rapid detection of allergic materials in foods. J. Food Hyg. Saf. 28: 124–129 (2013)
Prescott SL, Pawankar R, Allen KJ, Campbell DE, Sinn JK, Fiocchi A, Ebisawa M, Sampson HA, Beyer K, Lee BW. A global survey of changing patterns of food allergy burden in children. World Allergy Organ. J. 6: 21 (2013)
Reisch MP, Hochegger R, Stumr S, Korycanva K, Markl MC. Validation and comparison of two commercial ELISA kits and three in-house developed real-time PCR assays for the detection of potentially allergenic mustard in food. Food Chem. 174: 75–81 (2015)
Shang Y, Zhu P, Huang K, Liu W, Tian W, Luo Y, Xu W. A peach (Prunus persica)-specific gene, Lhcb2, used as an endogenous reference gene for qualitative and real-time quantitative PCR to detect fruit products. LWT-Food Sci. Technol. 55: 218–223 (2014)
Son DY, Yoon KR, Lee SI. Study of the most common allergic foods in Korea. Korean J. Food Sci. Technol. 34: 885–888 (2002)
Song JY, Kim JH, Kim HY. Detection of unapproved genetically modified potatoes in Korea using multiplex polymerase chain reaction. Food Control 80: 19–22 (2017)
Wen HW, Wlodzimierz BW, Thomas RD, Richard AD. Peanut allergy, peanut allergens, and methods for the detection of peanut contamination in food products. Compr. Rev. Food Sci. F. 6: 47–58 (2007)
Zhang C. Semi-nested multiplex PCR enhanced method sensitivity of species detection in further-processed meats. Food Control 31: 326–330 (2013)
Zhang W, Zhao Y, Xu Q, Chen Q. Development of a triplex real-time PCR assay for simultaneous detection of allergenic ingredients in processed food. Czech J. Food Sci. 36: 22–27 (2018)
Acknowledgements
This research was supported by Grant 16163MFDS003 from the Ministry of Food and Drug Safety in Korea.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Suh, SM., Park, SB., Kim, MJ. et al. Simultaneous detection of fruit allergen-coding genes in tomato, apple, peach and kiwi through multiplex PCR. Food Sci Biotechnol 28, 1593–1598 (2019). https://doi.org/10.1007/s10068-019-00591-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10068-019-00591-y