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Auto-microfluidic thin-film chip for genetically modified maize detection

Author:
Yun, Zhenyu, Peng, Liping, Huo, Shengnan, Wu, Xin, Zhou, Jun, Zhang, Hongguang, Li, Wenjing, Qi, Cai, Huang, Guangping
Source:
Food control 2017 v.80 pp. 360-365
ISSN:
0956-7135
Subject:
DNA, DNA fragmentation, DNA probes, alkaline phosphatase, color, corn, nucleic acid hybridization, nylon, polymerase chain reaction, screening, streptavidin, toxic substances, transgenic plants
Abstract:
As a thin-film chip method, reverse dot blot hybridization (RDBH) has been employed to detect hazardous substances, but an automatic RDBH instrument with low workload, high accuracy and stability is still urgently needed. This paper presents our newly-developed auto-microfluidic thin-film chip (AMTC) method for multiplex screening of genetically modified (GM) maize. With specific DNA probes for genetically modified (GM) maize being immobilized on a square nylon thin-film, it was placed into a micro-reaction cell of the AMTC device. Then biotin-labeled PCR products with target DNA fragments for template amplification were added to the micro-reaction cell using a microfluidic system. When the PCR products passed the square nylon thin-film, the target DNA fragments were captured by the complementary action of DNA, where the signal was visualized with streptavidin link-coupled alkaline phosphatase color development kit. The sensitivity of GM maize detection reached 0.1% quality percentage and its stability and consistency could satisfy the requirements for practical applications. Performance advantages of the ATMC are manifold, being embodied in aspects such as easy and straightforward operation, low costs and less workload.
Agid:
5710925