High-performance detection of Mycobacterium bovis in milk using digital LAMP

Yi Tao; Juanli Yun; Jian Wang; Peng Xu; Caiming Li; Hongtao Liu; Ying Lan; Jianzhang Pan; Wenbin Du

doi:10.1016/j.foodchem.2020.126945

High-performance detection of Mycobacterium bovis in milk using digital LAMP

Food Chem. 2020 Oct 15:327:126945. doi: 10.1016/j.foodchem.2020.126945. Epub 2020 May 6.

Authors

Yi Tao¹, Juanli Yun², Jian Wang², Peng Xu², Caiming Li¹, Hongtao Liu¹, Ying Lan³, Jianzhang Pan⁴, Wenbin Du⁵

Affiliations

¹ State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; College of Life Sciences, University of the Chinese Academy of Sciences, Beijing 10049, China.
² State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
³ State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China. Electronic address: [email protected].
⁴ Department of Chemistry, Zhejiang University, Hangzhou 310058, China. Electronic address: [email protected].
⁵ State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; College of Life Sciences, University of the Chinese Academy of Sciences, Beijing 10049, China; Savaid Medical School, University of the Chinese Academy of Sciences, Beijing 10049, China. Electronic address: [email protected].

PMID: 32447131
DOI: 10.1016/j.foodchem.2020.126945

Abstract

This paper described a high-performance molecular test for the detection of Mycobacterium bovis (M. bovis) based on digital loop-mediated isothermal amplification (dLAMP). M. bovis is a persistent pathogen that causes zoonotic tuberculosis and can infect both animals and human beings. The detection of M. bovis in milk samples is critical for effective control and prevention of zoonotic diseases but there lacks effective and sensitive methods. Here, we developed a convenient and low-cost system for M. bovis detection in milk, which incorporated automated DNA extraction and dLAMP by interfacial emulsification technique. Versus real-time PCR, dLAMP provides higher accuracy and sensitivity for direct M. bovis detection in milk, offering a limit of detection of 14 CFU/mL within 2 h. The dLAMP system can become a powerful platform for the detection of pathogens in complex samples and provide more reliable guidance for food safety testing, epidemiological research and clinical diagnosis.

Keywords: Digital loop-mediated isothermal amplification (dLAMP); Milk; Molecular detection; Mycobacterium bovis; Zoonotic tuberculosis.

MeSH terms

Animals
Milk / microbiology*
Mycobacterium bovis / isolation & purification*
Nucleic Acid Amplification Techniques
Real-Time Polymerase Chain Reaction