FOOD SCIENCE ›› 2019, Vol. 40 ›› Issue (8): 311-318.doi: 10.7506/spkx1002-6630-20180102-016

• Safety Detection • Previous Articles     Next Articles

Quantification of Genetically Modified Rice (Oryza sativa L.) Event KMD by Digital PCR Based on Droplet and Microfluidic Chip

DENG Tingting, HUANG Wensheng, GE Yiqiang, CHEN Ying   

  1. 1. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China; 2. Chinese Academy of Inspection and Quarantine, Beijing 100176, China; 3. China Rural Technology Development Center, Ministry of Science and Technology, Beijing 100045, China
  • Online:2019-04-25 Published:2019-05-05

Abstract: In this study, a quantitative duplex digital PCR method was established for insect-resistant genetically modified rice event KMD by using the sucrose phosphate synthase gene of rice and the KMD event-specific gene as the endogenous and exogenous genes, respectively. The optimal concentration of rice genomic DNA in the quantitative system was in the range of 10 pg to 13 ng, and the degree of template fracture and the annealing temperature of PCR amplification had little effect on the quantitative results. The absolute sensitivity and relative sensitivity of digital PCR were 1 copies/μL and 0.1%, respectively, while the relative sensitivity of the real-time PCR method established with the same pairs of primers and probes was only 1%. For standardization of the application of this method, an intra-laboratory repeatability validation was performed on two chip-based digital PCR platforms to verify the quantitative detection method established on the droplet digital PCR platform, and the results showed that the precision and accuracy of the assay still met the requirements for quantitative analysis. The method was proved to highly applicable the standardized quantitative analysis of GM rice KMD.

Key words: genetically modified rice, KMD, quantitative detection, digital PCR

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