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| Development of a Multiplex Liquidchip Assay for Rapid Identification of Fox and Mink Ingredients |
CHEN Ru1,**( ),DUAN Yan-yu1,GAO Xiao-bo2,LIU Zhi-ling1,YANG Jing1,MEI Ming-zhu1,TAN Xin3,LUO Hai-yan2 |
1 Technical Center, Guangzhou Customs District People’s Republic of China, Guangzhou 510623, China;
2 National Research Institute for Family Planning, Beijing 100081, China
3 School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, China |
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Abstract A novel nucleic acid detection method, based on the xMAP (flexible multi-analyte profiling) technology platform, was developed for high-throughput and simultaneous identification of fox and mink ingredients in products of animal origin. A duplex xMAP assay was established, the primers and probes were designed targeting a fox mitochondrial D-loop gene or a mink mitochondrial cytochrome b gene. The probes were incorporated with locked nucleic acids to improve detection efficiency. The assay accurately identified fox and mink DNA, without cross reactions with DNA samples of eighteen nontarget animal species. The LOD on purified fox and mink DNA was evaluated as 2.8pg/μl and 0.9pg/μl, respectively. The detection sensitivity on samples of experimental meat mixtures was demonstrated to be 0.05% (m/m). The assay successfully detected 32 mocked positive DNA sample of food and feeds, which each was made by adding fox DNA or mink DNA at 1% (V/V) proportion of the same concentration. In conclusion, the duplex xMAP assay provided rapid identification of fox and mink ingredients with high specificity and high sensitivity. The technique is suitable to be applied in food and feeds quality assurance systems and safety inspections.
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Received: 09 April 2019
Published: 17 December 2019
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Corresponding Authors:
Ru CHEN
E-mail: gd_chenr@sina.com
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