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Rapid and Sensitive Detection of sFAT-1 Transgenic Pigs by Visual Loop-Mediated Isothermal Amplification

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Abstract

Genetically modified (GM) livestock have the potential to contribute to improving the environment and human health, with consumption of fewer resources and reduced waste production. However, the transgene process also poses risks. The safety assessment and control of transgenic animal products have drawn wide attention, and the relevant regulations and technology are being developed. Quick testing technology plays a significant role in on-site and customs sampling. Nowadays, loop-mediated isothermal amplification (LAMP) was widely applied in nucleic acid analysis because of its simplicity, rapidity, high efficiency and specificity. In this study, a specific, sensitive detection system for detecting sFAT-1 transgenic pigs was designed. A set of six primers including two loop primers was designed for the target sequence. The DNA samples were amplified in less than 1 h at the optimized temperature and detecting by both Nephelometer LA-320c and unaided eyes directly adding calcein. The detection limit of sFAT-1 LAMP was as low as 1.26 ng/μL. Furthermore, blind tests of transgenic and non-transgenic DNA samples were all correctly detected. Hence, the results in this study demonstrated that LAMP is a very useful tool for transgenic detection.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (31501931), the 948 Program of China (2011-G11), and the Agricultural Science and Technology Innovation Program (ASTIP-IAS05).

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Authors and Affiliations

  1. Key Laboratory of Farm Animal Genetic Resources and Utilization & Inspection Test Center for Biosafety of Genetically Modified Animal and Feed, The Ministry of Agriculture of China (Beijing), Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100193, People’s Republic of China

    Chenyu Tao, Yalan Yang, Xunbi Li, Kui Li & Rong Zhou

  2. Department of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095, People’s Republic of China

    Chenyu Tao & Rong Zhou

  3. Hubei Key Laboratory of Animal Embryo Engineering and Molecular Breeding, Hubei Academy of Agricultural Sciences, Wuhan City, Hubei Province, China

    Xinmin Zheng & Hongyan Ren

Authors
  1. Chenyu Tao
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  2. Yalan Yang
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  3. Xunbi Li
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  4. Xinmin Zheng
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  5. Hongyan Ren
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  6. Kui Li
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  7. Rong Zhou
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Correspondence to Rong Zhou.

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Tao, C., Yang, Y., Li, X. et al. Rapid and Sensitive Detection of sFAT-1 Transgenic Pigs by Visual Loop-Mediated Isothermal Amplification. Appl Biochem Biotechnol 179, 938–946 (2016). https://doi.org/10.1007/s12010-016-2041-8

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  • DOI: https://doi.org/10.1007/s12010-016-2041-8

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