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

Jinhua Liu,Xinmin Zhang,Yanyu Shi,Lianpeng Wu,Siying Teng

doi:10.1155/2017/9319035

Jinhua Liu, Xinmin Zhang + Show 3 more

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https://doi.org/10.1155/2017/9319035

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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.

Highlights

Raccoon dog is a member of the canid family; its fur is an economically important product
The felt made from the fur of raccoon dogs is a highly expensive object in the commercial market; raccoon dog meat is much cheaper than other kinds of meat due to its unpleasant taste and poor texture
Raccoon dog meat is often included as a subsidiary component in compound meat mixtures that are primarily made from mutton, beef, pork, and so forth

Summary

Introduction

Raccoon dog is a member of the canid family; its fur is an economically important product. The felt made from the fur of raccoon dogs is a highly expensive object in the commercial market; raccoon dog meat is much cheaper than other kinds of meat due to its unpleasant taste and poor texture. When it is included in the composition of meat mixtures, especially in dressed meat, its appearance and taste resemble the meat obtained from other animals. Raccoon dog meat is often included as a subsidiary component in compound meat mixtures that are primarily made from mutton, beef, pork, and so forth.

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