Validation of end-point and real-time PCR methods for the rapid detection of soy allergen in processed products

This work describes the development and validation of two PCR methods, end-point and real-time PCR, for the detection of soy protein in a wide rage of foodstuffs. These techniques are reliable and sensitive, allowing detection of trace amounts of soybean in processed products. TaqMan real-time PCR was the simpler and more rapid process, with a higher potential for automation and, therefore, currently the most suitable screening method. To verify correct operation of the proposed methodology, ELISA was used for quantitative determination of soy protein. In addition, 35 meat, fish and bakery processed products, which could potentially contain soy but was not declared on the label, were tested for the presence of soy DNA using the proposed methods. The methodologies will be valuable in issues regarding the presence of soy protein in processed products, especially in verifying labelling and security regulations to protect consumer's rights.

[1]  M. Abramson,et al.  Reported adverse food reactions overestimate true food allergy in the community , 2002, European Journal of Clinical Nutrition.

[2]  E Anklam,et al.  Methods for allergen analysis in food: a review , 2004, Food additives and contaminants.

[3]  C Y Ou,et al.  Polymerase chain reaction. , 1988, The Journal of infectious diseases.

[4]  S. Radu,et al.  Detection of genetically modified soy in processed foods sold commercially in Malaysia by PCR-based method , 2006 .

[5]  J. Thompson,et al.  The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. , 1997, Nucleic acids research.

[6]  M. Marina,et al.  Analysis of Soyabean Proteins in Meat Products: A Review , 2002, Critical reviews in food science and nutrition.

[7]  Rolf Meyer,et al.  Polymerase chain reaction (PCR) in the quality and safety assurance of food: Detection of soya in processed meat products , 1996, Zeitschrift fur Lebensmittel-Untersuchung und -Forschung.

[8]  A. Burks,et al.  Peanut and soy allergy: a clinical and therapeutic dilemma , 2000, Allergy.

[9]  T. Kuchta,et al.  Comparison of three types of methods for the isolation of DNA from flours, biscuits and instant paps , 2004 .

[10]  M. Shimokomaki,et al.  Textured Soy Protein Quantification in Commercial Hamburger , 2001 .

[11]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[12]  S. Rogers,et al.  Extraction of DNA from plant tissues , 1989 .

[13]  H. Matsufuji,et al.  Extraction and detection of endogenous soybean DNA from fermented foods , 2006 .

[14]  Kathy Messens,et al.  Evaluation and optimisation of five different extraction methods for soy DNA in chocolate and biscuits. Extraction of DNA as a first step in GMO analysis , 2004 .

[15]  J. Lüthy,et al.  Detection of wheat adulteration of spelt flour and products by PCR , 2001 .

[16]  Isabel Mafra,et al.  Comparative study of DNA extraction methods for soybean derived food products , 2008 .

[17]  B. Lu,et al.  Rapid and reliable identification of rice genomes by RFLP analysis of PCR-amplified Adh genes. , 2001, Genome.

[18]  J. G. Endres Soy Protein Products: Characteristics, Nutritional Aspects, and Utilization , 2001 .

[19]  K. Kamata,et al.  Applicability of quantitative PCR to soy processed foods containing Roundup Ready Soy , 2007 .

[20]  Montserrat Espiñeira,et al.  Genetic identification of squids (families Ommastrephidae and Loliginidae) by PCR-RFLP and FINS methodologies. , 2007, Journal of agricultural and food chemistry.

[21]  Xuejun Gao,et al.  Detection of Roundup Ready soy in highly processed products by triplex nested PCR , 2007 .

[22]  A. C. Arisi,et al.  Recombinant DNA in meat additives: Specific detection of Roundup Ready™ soybean by nested PCR , 2007 .

[23]  I. Malmheden Yman,et al.  A study on severe food reactions in Sweden – is soy protein an underestimated cause of food anaphylaxis? , 1999, Allergy.

[24]  E. Wit Design and Analysis of DNA Microarray Investigations , 2004, Human Genomics.

[25]  T. A. Hall,et al.  BIOEDIT: A USER-FRIENDLY BIOLOGICAL SEQUENCE ALIGNMENT EDITOR AND ANALYSIS PROGRAM FOR WINDOWS 95/98/ NT , 1999 .