title: A highly sensitive and specific method for the screening detection of genetically modified organisms based on digital PCR without pretreatment.
abstract: Digital PCR has developed rapidly since it was first reported in the 1990 s. It was recently reported that an improved method facilitated the detection of genetically modified organisms (GMOs). However, to use this improved method, the samples must be pretreated, which could introduce inaccuracy into the results. In our study, we explored a pretreatment-free digital PCR detection method for the screening for GMOs. We chose the CaMV35s promoter and the NOS terminator as the templates in our assay. To determine the specificity of our method, 9 events of GMOs were collected, including MON810, MON863, TC1507, MIR604, MIR162, GA21, T25, NK603 and Bt176. Moreover, the sensitivity, intra-laboratory and inter-laboratory reproducibility of our detection method were assessed. The results showed that the limit of detection of our method was 0.1%, which was lower than the labeling threshold level of the EU. The specificity and stability among the 9 events were consistent, respectively. The intra-laboratory and inter-laboratory reproducibility were both good. Finally, the perfect fitness for the detection of eight double-blind samples indicated the good practicability of our method. In conclusion, the method in our study would allow more sensitive, specific and stable screening detection of the GMO content of international trading products.
authors.affiliation: The Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing, 100029, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Fu
authors.affiliation: Laboratory of Food Safety, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
authors.forename: Pengyu
authors.initials: P
authors.lastname: Zhu
authors.affiliation: Laboratory of Food Safety, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
authors.forename: Chenguang
authors.initials: C
authors.lastname: Wang
authors.affiliation: 1] Laboratory of Food Safety, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China [2] The Supervision, Inspection and Testing Center of Genetically Modified Organisms, Ministry of Agriculture, Beijing, 100083, China.
authors.forename: Kunlun
authors.initials: K
authors.lastname: Huang
authors.affiliation: Guangxi Entry-Exit Inspection and Quarantine Bureau, Guangxi, 530028, China.
authors.forename: Zhixin
authors.initials: Z
authors.lastname: Du
authors.affiliation: 1] Laboratory of Food Safety, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China [2] The Supervision, Inspection and Testing Center of Genetically Modified Organisms, Ministry of Agriculture, Beijing, 100083, China.
authors.forename: Wenying
authors.initials: W
authors.lastname: Tian
authors.affiliation: The Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing, 100029, China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Wang
authors.affiliation: The Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing, 100029, China.
authors.forename: Huiyu
authors.initials: H
authors.lastname: Wang
authors.affiliation: 1] Laboratory of Food Safety, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China [2] The Supervision, Inspection and Testing Center of Genetically Modified Organisms, Ministry of Agriculture, Beijing, 100083, China.
authors.forename: Wentao
authors.initials: W
authors.lastname: Xu
authors.affiliation: The Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing, 100029, China.
authors.forename: Shuifang
authors.initials: S
authors.lastname: Zhu
mesh_terms: D018744:DNA, Plant / Q000235:genetics*; D018628:Gene Dosage; D057230:Limit of Detection; D015588:Observer Variation; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D011401:Promoter Regions, Genetic; D015203:Reproducibility of Results; D012815:Signal Processing, Computer-Assisted; D019076:Transgenes; D003313:Zea mays / Q000235:genetics*
chemical_list: D018744:DNA, Plant