Development of a triplex real-time PCR for simultaneous detection of allergenic ingredients in processed food
Wenju Zhang, Yulei Zhao, Qingjin Xu, Qin Chenhttps://doi.org/10.17221/28/2017-CJFSCitation:Zhang W., Zhao Y., Xu Q., Chen Q. (2018): Development of a triplex real-time PCR for simultaneous detection of allergenic ingredients in processed food. Czech J. Food Sci., 36: 22-27.
SYBR Green real-time or quantitative PCR (Q-PCR) is a suitable system in which to establish a multiplex method to detect allergenic ingredients in food. In this study, a triplex Q-PCR method was developed to detect trace amounts of peanut, soybean and sesame in processed food. Specific PCR primer sets were designed and the concentration of the primers used in the triplex PCR was optimised. The triplex method showed high specificity and sensitivity which were similar to those of the simplex method, and it was applied for the detection of allergenic ingredients in commercially available processed food. The results demonstrate that the developed triplex Q-PCR is a quick, reliable and efficient method for the detection of allergenic ingredients in processed food.Keywords:
peanut; sesame; soybean; SYBR Green; triplex Q-PCRReferences:
Agrimonti Caterina, Pirondini Andrea, Marmiroli Marta, Marmiroli Nelson (2015): A quadruplex PCR (qxPCR) assay for adulteration in dairy products. Food Chemistry, 187, 58-64 https://doi.org/10.1016/j.foodchem.2015.04.017BOUSQUET J, LOCKEY R, MALLING H (1998): Allergen immunotherapy: Therapeutic vaccines for allergic diseases A WHO position paper☆☆☆★★★♢. Journal of Allergy and Clinical Immunology, 102, 558-562 https://doi.org/10.1016/S0091-6749(98)70271-4Davis P.J., Smales C.M., James D.C. (2001): How can thermal processing modify the antigenicity of proteins? Allergy, 56: 56–60.Hernández Marta, Esteve Teresa, Pla Maria (2005): Real-Time Polymerase Chain Reaction Based Assays for Quantitative Detection of Barley, Rice, Sunflower, and Wheat. Journal of Agricultural and Food Chemistry, 53, 7003-7009 https://doi.org/10.1021/jf050797jHerrero Beatriz, Vieites Juan M., Espiñeira Montserrat (2014): Development of an in-house fast real-time PCR method for detection of fish allergen in foods and comparison with a commercial kit. Food Chemistry, 151, 415-420 https://doi.org/10.1016/j.foodchem.2013.11.042Hird H., Lloyd J., Goodier R., Brown J., Reece P. (2003): Detection of peanut using real-time polymerase chain reaction. European Food Research and Technology, 217, 265-268 https://doi.org/10.1007/s00217-003-0726-zNavarro E., Serrano-Heras G., Castaño M.J., Solera J. (2015): Real-time PCR detection chemistry. Clinica Chimica Acta, 439, 231-250 https://doi.org/10.1016/j.cca.2014.10.017Pafundo Simona, Gullì Mariolina, Marmiroli Nelson (2010): Multiplex real-time PCR using SYBR® GreenER™ for the detection of DNA allergens in food. Analytical and Bioanalytical Chemistry, 396, 1831-1839 https://doi.org/10.1007/s00216-009-3419-zPalle-Reisch Monika, Hochegger Rupert, Cichna-Markl Margit (2015): Development and validation of a triplex real-time PCR assay for the simultaneous detection of three mustard species and three celery varieties in food. Food Chemistry, 184, 46-56 https://doi.org/10.1016/j.foodchem.2015.03.049Poms R.E., Klein C.L., Anklam E. (2004): Methods for allergen analysis in food: a review. Food Additives & Contaminants, 21: 1–31.Rastogi N. K., Raghavarao K. S. M. S., Balasubramaniam V. M., Niranjan K., Knorr D. (2007): Opportunities and Challenges in High Pressure Processing of Foods. Critical Reviews in Food Science and Nutrition, 47, 69-112 https://doi.org/10.1080/10408390600626420REDMOND ELIZABETH C., GRIFFITH CHRISTOPHER J. (2003): Consumer Food Handling in the Home: A Review of Food Safety Studies. Journal of Food Protection, 66, 130-161 https://doi.org/10.4315/0362-028X-66.1.130Reed Gudrun H, Kent Jana O, Wittwer Carl T (2007): High-resolution DNA melting analysis for simple and efficient molecular diagnostics. Pharmacogenomics, 8, 597-608 https://doi.org/10.2217/146224126.96.36.1997Shin Sang Phil, Ishitani Hiroe, Shirakashi Sho (2016): Development of a multiplex PCR to detect Kudoa spp. and to distinguish Kudoa septempunctata in olive flounder Paralichthys olivaceus. Aquaculture, 464, 37-41 https://doi.org/10.1016/j.aquaculture.2016.06.005Sicherer Scott H., Sampson Hugh A. (2007): Peanut allergy: Emerging concepts and approaches for an apparent epidemic. Journal of Allergy and Clinical Immunology, 120, 491-503 https://doi.org/10.1016/j.jaci.2007.07.015Sicherer Scott H., Sampson Hugh A. (2014): Food allergy: Epidemiology, pathogenesis, diagnosis, and treatment. Journal of Allergy and Clinical Immunology, 133, 291-307.e5 https://doi.org/10.1016/j.jaci.2013.11.020Wei Shuang, Zhao Hui, Xian Yuyin, Hussain Malik A., Wu Xiyang (2014): Multiplex PCR assays for the detection of Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio vulnificus, and Vibrio cholerae with an internal amplification control. Diagnostic Microbiology and Infectious Disease, 79, 115-118 https://doi.org/10.1016/j.diagmicrobio.2014.03.012Xu Yi-Gang, Sun Liu-Mei, Wang Yu-Sai, Chen Pei-Pei, Liu Zhong-Mei, Li Yi-Jing, Tang Li-Jie (2017): Simultaneous detection of Vibrio cholerae , Vibrio alginolyticus , Vibrio parahaemolyticus and Vibrio vulnificus in seafood using dual priming oligonucleotide (DPO) system-based multiplex PCR assay. Food Control, 71, 64-70 https://doi.org/10.1016/j.foodcont.2016.06.024Zhang Wen-Ju, Cai Qin, Guan Xiao, Chen Qin (2015): Detection of peanut (Arachis hypogaea) allergen by Real-time PCR method with internal amplification control. Food Chemistry, 174, 547-552 https://doi.org/10.1016/j.foodchem.2014.11.091