Real-time PCR multiplex method for the quantification of Roundup Ready soybean in raw material and processed food |
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Authors: | Nicoletta Foti Roberta Onori Erica Donnarumma Barbara De Santis Marina Miraglia |
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Affiliation: | (1) National Institute of Health, National Center for Food Quality and Risk Assessment, GMO and Mycotoxins Unit, viale Regina Elena 299, 00161 Rome, Italy;(2) Present address: JRC Institute for Health and Consumer Protection, via Enrico Fermi 1, 21020 Ispra, Varese, Italy |
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Abstract: | This paper describes a quantitative real-time multiplex PCR method optimised for the ABI PRISM® 7700 Sequence Detection System (SDS) and TaqMan® chemistry for Roundup Ready® 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 (CRM IRMM-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. |
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Keywords: | Genetically modified organisms Quantification multiplex Real-time PCR Roundup? Ready soybean? Processed food |
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