Interlaboratory trial validation of an event-specific qualitative polymerase chain reaction-based detection method for genetically modified RT73 rapeseed

• Published in: Journal of AOAC International Vol. 90; no. 6; pp. 1639 - 1646
• Main Authors: Pan, L, Zhang, S, Yang, L, Broll, H, Tian, F, Zhang, D
• Format: Journal Article
• Language: English
• Published: Gaithersburg, MD AOAC International 01.11.2007; Oxford University Press
• Subjects: Biological and medical sciences; Brassica napus var. napus; Brassica rapa - chemistry; Brassica rapa - genetics; canola; cell lines; cooperative research; detection; DNA Primers; DNA, Plant - genetics; DNA, Plant - isolation & purification; Fish and seafood industries; Food industries; Fundamental and applied biological sciences. Psychology; gene banks; General aspects; Genetic aspects; genetically modified foods; Genetically modified plants; laboratories; Methods; Methods of analysis, processing and quality control, regulation, standards; molecular sequence data; Plants, Genetically Modified - chemistry; Plants, Genetically Modified - genetics; Polymerase chain reaction; product authenticity; Properties; qualitative analysis; Rapeseed; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction; transgenes; transgenic plants; China; Bass; Polymerase chain reaction; Validation; Edible fish; Oilseed; Analysis method; Rapeseed; Control method; Molecular biology; Detection; Genetically modified organism
• ISSN: 1060-3271; 1944-7922
• DOI: 10.1093/jaoac/90.6.1639

The qualitative event-specific polymerase chain reaction detection method of genetically modified (GM) RT73 rapeseed was developed based on the cloned 3' end flanking sequence of RT73 rapeseed integration. The specificity of the method for GM RT73 rapeseed was validated using several different GM rapeseed lines, GM maize lines, GM soybean line, non-GM rapeseed, and other non-GM crops. In this study, the developed method was validated through an interlaboratory study by 12 laboratories from 6 countries. The sensitivity of this method was evaluated using several mixed rapeseed meals with different GM RT73 rapeseed contents from 5.0 to 0.01% prepared by our laboratory. The evaluated results showed that all of the rapeseed endogenous reference high mobility group protein gene (HMG I/Y), figwort mosaic virus 35S (FMV 35S) promoter, and RT73 event-specific fragment could be detected from rapeseed samples at 0.1% (w/w) with a confidence level of more than 95%. All results from the 12 laboratories indicated that the developed method could be considered fit for the detection and identification of GM RT73 rapeseed.