title: Developing a matrix reference material for screening of transgenic rice.
abstract: Certified reference materials (CRMs) that are compatible with detection methods are needed to detect genetically modified organisms (GMOs). Screening is the first detection step in determining the possible presence of GMO ingredients in food or feed; however, screening has been hindered by the lack of GMO CRMs. In this study, transgenic rice materials were developed via the transformation of a construct harboring 11 commonly used screening elements. Digital PCR was utilized to identify a homozygous single-copy line termed SDrice. The qualitative detections of 11 elements in 21 transgenic materials demonstrated that the genomic DNA of the SDrice was suitable for use as a positive control in the screening of GMO ingredients. The suitability of SDrice as reference material was further checked by testing the sensitivity of 11 known conventional PCR assays, ranging from 10 to 50 copies of the SDrice genome. The standard curves that were created using SDrice DNA series as calibrators all exhibited good linearities in the relationships of the Ct values with the template copy numbers in these 11 real-time PCR assays. The LODs of the real-time PCR assays were estimated to be two to five copies of the SDrice genome. Comparisons of the SDrice with other GM rice revealed that significant differences existed in both the intercepts of the standard curves and the ΔCt values of the exogenous and reference genes for the P-35S, T-nos, HPT, T-35S, and Bar assays; the SDrice was not fit for quantification of other GM rice events. This study provided a matrix reference material (RM) that was suitable for screening GM rice, determination of sensitivity and a LOD of PCR assays, and overcame some of the drawbacks of plasmid DNA as reference material.
authors.affiliation: Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan, 430062, China.|Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan, 430062, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan, 430062, China.|Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan, 430062, China.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan, 430062, China.|Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan, 430062, China.
authors.forename: Xiaofei
authors.initials: X
authors.lastname: Li
authors.affiliation: Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan, 430062, China.|Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan, 430062, China.
authors.forename: Yulei
authors.initials: Y
authors.lastname: Wang
authors.affiliation: College of Life Science, South-Central University for Nationalities, 182 Min-Yuan Road, 430074, Wuhan, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan, 430062, China.|Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan, 430062, China.
authors.forename: Yunjing
authors.initials: Y
authors.lastname: Li
authors.affiliation: Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan, 430062, China. wugang@caas.cn.|Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan, 430062, China. wugang@caas.cn.
authors.forename: Gang
authors.initials: G
authors.lastname: Wu
mesh_terms: D018744:DNA, Plant / Q000235:genetics; D030841:Food, Genetically Modified; D012275:Oryza / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
chemical_list: D018744:DNA, Plant
keywords: Comparability; Matrix RM; Positive control; Screening; Transgenic rice