title: Development and validation of a 48-target analytical method for high-throughput monitoring of genetically modified organisms.
abstract: The rapid increase in the number of genetically modified (GM) varieties has led to a demand for high-throughput methods to detect genetically modified organisms (GMOs). We describe a new dynamic array-based high throughput method to simultaneously detect 48 targets in 48 samples on a Fludigm system. The test targets included species-specific genes, common screening elements, most of the Chinese-approved GM events, and several unapproved events. The 48 TaqMan assays successfully amplified products from both single-event samples and complex samples with a GMO DNA amount of 0.05 ng, and displayed high specificity. To improve the sensitivity of detection, a preamplification step for 48 pooled targets was added to enrich the amount of template before performing dynamic chip assays. This dynamic chip-based method allowed the synchronous high-throughput detection of multiple targets in multiple samples. Thus, it represents an efficient, qualitative method for GMO multi-detection.
authors.affiliation: 1] Key Laboratory of Oil Crop Biology, Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, People's Republic of China [2] Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan 430062, People's Republic of China.
authors.forename: Xiaofei
authors.initials: X
authors.lastname: Li
authors.affiliation: 1] Key Laboratory of Oil Crop Biology, Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, People's Republic of China [2] Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan 430062, People's Republic of China.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Wu
authors.affiliation: 1] Key Laboratory of Oil Crop Biology, Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, People's Republic of China [2] Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan 430062, People's Republic of China.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.affiliation: 1] Key Laboratory of Oil Crop Biology, Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, People's Republic of China [2] Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan 430062, People's Republic of China.
authors.forename: Yunjing
authors.initials: Y
authors.lastname: Li
authors.affiliation: Agro-Biotechnology Research Institute, Jilin Academy of Agricultural Sciences, No.1363 Caiyu Avenue, Changchun 130033, People's Republic of China.
authors.forename: Likun
authors.initials: L
authors.lastname: Long
authors.affiliation: Agro-Biotechnology Research Institute, Jilin Academy of Agricultural Sciences, No.1363 Caiyu Avenue, Changchun 130033, People's Republic of China.
authors.forename: Feiwu
authors.initials: F
authors.lastname: Li
authors.affiliation: 1] Key Laboratory of Oil Crop Biology, Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, People's Republic of China [2] Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan 430062, People's Republic of China.
authors.forename: Gang
authors.initials: G
authors.lastname: Wu
mesh_terms: D018744:DNA, Plant / Q000032:analysis* / Q000302:isolation & purification; D057166:High-Throughput Screening Assays; D020411:Oligonucleotide Array Sequence Analysis; D030821:Plants, Genetically Modified / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D013045:Species Specificity
chemical_list: D018744:DNA, Plant