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An integrated digital polymerase chain reaction chip for multiplexed meat adulteration detection
Authors:Chengzhuang Yu  Shijie Dai  Ziqi Zhang  Shanshan Li  Jingmeng Cheng  Hezhi Hu  Jayne Wu  Junwei Li
Institution:1. Hebei Key Laboratory of Smart Sensing and Human-robot Interactions, School of Mechanical Engineering, Hebei University of Technology, Tianjin, P. R. China;2. State Key Laboratory of Reliability and Intelligence of Electrical Equipment, Hebei University of Technology, Tianjin, P. R. China;3. Department of Electrical Engineering and Computer Science, The University of Tennessee, Knoxville, Tennessee, USA;4. Institute of Biophysics, School of Health Sciences and Biomedical Engineering, Hebei University of Technology, Tianjin, P. R. China
Abstract:Meat adulteration detection is a common concern of consumers. Here, we proposed a multiplex digital polymerase chain reaction method and a low-cost device for meat adulteration detection. Using a polydimethylsiloxane microfluidic device, polymerase chain reaction reagents could be pump-free loaded into microchambers (40 × 40 chambers) automatically. Due to the independence of multiplex fluorescence channels, deoxyribonucleic acid templates extracted from different animal species could be distinguished by one test. In this paper, we designed primers and probes for four types of meat (beef, chicken, pork, and duck) and labeled each of the four fluorescent markers (hexachlorocyclohexane HEX], 6-carboxyfluorescein FAM], X-rhodamine ROX], and cyanine dyes 5 CY5]) on the probes. Specific detection and mixed detection experiments were performed on four types of meat, realizing a limit of detection of 3 copies/µL. A mixture of four different species can be detected by four independent fluorescence channels. The quantitative capability of this method is found to meet the requirements of meat adulteration detections. This method has great potential for point-of-care testing together with portable microscopy equipment.
Keywords:degassed  digital PCR  meat authentication  multiplex  polymerase chain reaction
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