Comparison of the sensitivity of determining soyeabean allergens by ELISA method and SYBR green I

Abstract

The aim of this study was to compare the suitability of two methods for detecting defatted soybean powder; SYBR Green I Real-time PCR and enzyme-linked immunosorbent assay (ELISA). Analysis of 20 artificially contaminated samples prepared by simple dilution with wheat flour revealed that both techniques were able to detect defatted soybean powder, although there were significant differences between the two methods. Wheat flour contamination with defatted soybean powder was detected in samples 1-5, (0.012 %; 120 mg.kg-1), but not in samples with lower contamination with soybean powder saples 6-20 using SYBR Green I real-time PCR. Samples 1-10 could not be quantified by ELISA as the absorbance values were greater than the detection limit, and while samples 11-20 were measured, only the values of samples 16, 17 and 18 were within the guaranteed quantification range specified by the ELISA kit manufacturer. Defatted soybean powder contamination was detected in samples 19 and 20, but absorbance values were highly similar to those of the negative control sample.

Keywords

• ELISA
• PCR
• soybean
• allergen
• SYBR Green I