title: A novel quadruplex real-time PCR method for simultaneous detection of Cry2Ae and two genetically modified cotton events (GHB119 and T304-40).
abstract: BACKGROUND
  To date, over 150 genetically modified (GM) crops are widely cultivated. To comply with regulations developed for genetically modified organisms (GMOs), including labeling policies, many detection methods for GMO identification and quantification have been developed.
  
  
  RESULTS
  To detect the entrance and exit of unauthorized GM crop events in China, we developed a novel quadruplex real-time PCR method for simultaneous detection and quantification of GM cotton events GHB119 and T304-40 in cotton-derived products (based on the 5'-flanking sequence) and the insect-resistance gene Cry2Ae. The limit of detection was 10 copies for GHB119 and Cry2Ae and 25 copies for T304-40. The limit of quantification was 25 copies for GHB119 and Cry2Ae and 50 copies for T304-40. Moreover, low bias and acceptable standard deviation and relative standard deviation values were obtained in quantification analysis of six blind samples containing different GHB119 and T304-40 ingredients.
  
  
  CONCLUSIONS
  The developed quadruplex quantitative method could be used for quantitative detection of two GM cotton events (GHB119 and T304-40) and Cry2Ae gene ingredient in cotton derived products.
authors.affiliation: GMO Detection Laboratory, Shanghai Entry-Exit Inspection and Quarantine Bureau, 1208 Minsheng Road, Shanghai 200135, PR China. idealne@163.com.
authors.forename: Xiang
authors.initials: X
authors.lastname: Li
authors.forename: Xiuxiu
authors.initials: X
authors.lastname: Wang
authors.forename: Jielin
authors.initials: J
authors.lastname: Yang
authors.forename: Yueming
authors.initials: Y
authors.lastname: Liu
authors.forename: Yuping
authors.initials: Y
authors.lastname: He
authors.forename: Liangwen
authors.initials: L
authors.lastname: Pan
mesh_terms: D001483:Base Sequence; D056931:Cryptochromes / Q000235:genetics*; D018744:DNA, Plant / Q000032:analysis*; D003368:Gossypium / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D016415:Sequence Alignment
chemical_list: D056931:Cryptochromes; D018744:DNA, Plant; D010940:Plant Proteins