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Journal of Food Quality
Volume 2017, Article ID 9319035, 8 pages
https://doi.org/10.1155/2017/9319035
Research Article

Establishment and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Detection of Raccoon Dog in Meat Mixtures

1School of Pharmaceutical Sciences, Jilin University, Changchun, Jilin, China
2Jilin Entry-Exit Inspection and Quarantine Bureau, Changchun, Jilin, China
3Jilin Product Quality Supervision Inspection, Changchun, Jilin, China
4Department of Ophthalmology, First Hospital of Jilin University, Changchun, Jilin, China

Correspondence should be addressed to Xinmin Zhang; nc.ude.ulj@nimnixgnahz

Received 23 August 2016; Revised 21 November 2016; Accepted 19 December 2016; Published 10 January 2017

Academic Editor: Alejandro Hernández

Copyright © 2017 Jinhua Liu et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Raccoon dog (Nyctereutes procyonoides) is an economically important animal used for fur production, but consuming its meat is injurious to human health. Currently, no rapid and sensitive method for detecting raccoon dog meat in meat mixtures is available. In this study, we developed an easily applicable, rapid, and economically feasible method for identifying the presence of raccoon dog in meat mixtures based on loop-mediated isothermal amplification (LAMP). Four sets of LAMP primers were tested at different temperatures, and the primers that worked best at 62°C (set 2) were determined. In the LAMP assay, there was no cross-reactivity with the meat procured from other species of animals and the detection limit of DNA concentration was 0.1 pg·μL−1, slightly higher than TaqMan real-time PCR (0.01 pg·μL−1), but sensitivity of 0.1 pg·μL−1 complies with most requirements of routine analysis. Moreover, by the LAMP method, the meat mixtures containing more than 0.5% of the raccoon dog component were directly detected (without DNA extraction) in the supernatant isolated from the meat mixtures after performing repeated cycles of thawing and freezing of minced meat mixtures. Our results show that LAMP assay is a valuable, straightforward, and sensitive detection tool for identification of raccoon dog meat in mixtures.