Real-time PCR multiplex method for the quantification of Roundup Ready soybean in raw material and processed food

摘要

This paper describes a quantitative real-time multiplex PCR method optimised for the ABI PRISM~R 7700 Sequence Detection System (SDS) and TaqMan~R chemistry for Roundup Ready~R Soybean (RRS) in raw material and processed food. This method has the advantage of performing the amplification of the target taxon lectin gene and the genetically modified (GM) target sequence in the same test tube. The quantification is based on a calibration curve obtained with the DNA extracted from Certified Reference Material standards (CRMIRMM-410: R) in the range 0.1-5% RRS. The method was validated in-house by using CRMs and the applicability was verified on three mixtures of soybean flour at 1, 5, 10% of RRS respectively and on prepared baked products (biscuits and plum cakes). The statistical parameters of the method were found to be satisfactory both for soybean flour and baked products. In the process the absolute LOD and LOQ was 7.1 and 35.5 copy number respectively; the relative LOD and LOQ was 0.03 and 0.01% of RRS respectively.