Citation: XU Jun-Yi, CAO Ji-Juan, ZHENG Qiu-Yue, YANG Li-Li, WANG Yong, ZOU Ming-Qiang. High-throughput Identification and Detection of 17 Transgenic Maize Events Based on Taqman Microfluidic Chip Technology[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(11): 1477-1485. doi: 10.19756/j.issn.0253-3820.201287 shu

High-throughput Identification and Detection of 17 Transgenic Maize Events Based on Taqman Microfluidic Chip Technology

1.
Technology Center of Dalian Customs District, Dalian 116001, China;
2.
Key Laboratory of Biotechnology and Bioresources Utilization of Ministry of Education, Dalian Minzu University, Dalian 116600, China;
3.
Tianjin Academy of Agricultural Sciences, Tianjin 300192, China;
4.
Chinese Academy of Inspection and Quarantine, Beijing 100123, China;
5.
China Inspection Laboratory Technologies Co. Ltd., Beijing 100123, China

Corresponding author: CAO Ji-Juan, caojijuan@dlnu.edu.cn
Received Date: 9 May 2020
Revised Date: 1 September 2020

Fund Project: This work was supported by the National Special Project on Breeding New Varieties of Genetically Modified Organisms(No. 2018ZX08012-001).

Taqman microfluidic chip technology was applied to the high-throughput identification and detection of 17 transgenic maize events on a real-time fluorescent PCR platform. In one PCR amplification process, 2 maize endogenous genes (hmgA gene, adh1 gene) and 1 loading control gene (18S gene) were used as internal reference, which could simultaneously complete totally 17 events of transgenic maize single-well single-plex amplification parallel detection, including TC1507, NK603, MON87640, MON863, MON810, MIR162, GA21, DAS40278, BT176, BT11, 98140, 59122, 3272, MON89034, MIR604, MON88017 and T25, and the limit of detection reached 10-20 copies. The method showed high specificity and sensitivity, and the detection result obtained by this method was completely consistent with that of the single real-time fluorescent PCR method of "gold standard". The method had the advantages of parallel detection of multiple samples and multiple targets, and provided an efficient and rapid approach for the identification of mixture of multiple events of genetically modified products. With this method, 9 batches of transgenic maize samples were detected from 16 batches of imported practical maize, containing 1-8 events. This method could be used for high-throughput detection of mixed transgenic multi-events in imported agricultural products at the port.
- Taqman microfluidic chip,
- Transgenic maize,
- Event identification,
- Multi-events parallel detection
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