Rapid Detection of Salmonella and Listeria monocytogenes in Fresh Beef Using Loop-Mediated Isothermal Amplification

被引:0
作者
Wang, Xiaoran [1 ]
Jia, Kaixiang [1 ]
Ran, Jinrong [1 ]
Du, Yangyang [1 ]
Yu, Xiaoying [1 ]
Cao, Xuefeng [1 ]
Peng, Lianci [1 ]
Chen, Weifeng [2 ]
Fang, Rendong [1 ]
Li, Zhiwei [1 ]
机构
[1] Southwest Univ, Coll Vet Med, Joint Int Res Lab Anim Hlth & Anim Food Safety, Chongqing 400715, Peoples R China
[2] Henan Univ Anim Husb & Econ, Sch Food & Bioengn, Zhengzhou, Peoples R China
关键词
loop-mediated isothermal amplification (LAMP); Salmonella; Listeria monocytogenes; rapid detection; fresh beef; LAMP; ENTERICA; CHAIN; FOOD;
D O I
10.1089/fpd.2025.0018
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
This study developed and optimized loop-mediated isothermal amplification (LAMP) assays for rapid detection of Salmonella and L. monocytogenes in fresh beef. LAMP primers targeting the invA gene of Salmonella and hly gene of L. monocytogenes were used. Reaction conditions including temperature, dNTP concentration, Mg2+ concentration, primer ratio, and Bst DNA polymerase amount were optimized for each pathogen. Differences were observed between the two pathogens in optimal temperature, Mg2+ concentration, and Bst DNA polymerase requirements, highlighting the importance of pathogen-specific optimization. The optimized assays demonstrated high sensitivity with detection limits of 120 fg/mu L for Salmonella and 130 fg/mu L for L. monocytogenes, achieving detection within 40 and 60 min, respectively. Specificity tests confirmed both assays were highly specific for their target pathogens in fresh beef samples with no cross-reactivity observed. Addition of hydroxynaphthol blue enabled simple visual detection of positive results, eliminating the need for specialized equipment. The developed LAMP assays offer rapid, sensitive, and specific detection of these important foodborne pathogens.
引用
收藏
页数:8
相关论文
共 29 条
[21]   Trends and advances in food analysis by real-time polymerase chain reaction [J].
Salihah, Nur Thaqifah ;
Hossain, Mohammad Mosharraf ;
Lubis, Hamadah ;
Ahmed, Minhaz Uddin .
JOURNAL OF FOOD SCIENCE AND TECHNOLOGY-MYSORE, 2016, 53 (05) :2196-2209
[22]   Loop-Mediated Isothermal Amplification: From Theory to Practice [J].
Shirshikov, F. V. ;
Bespyatykh, J. A. .
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY, 2022, 48 (06) :1159-1174
[23]   The Listeria transcriptional landscape from saprophytism to virulence [J].
Toledo-Arana, Alejandro ;
Dussurget, Olivier ;
Nikitas, Georgios ;
Sesto, Nina ;
Guet-Revillet, Helene ;
Balestrino, Damien ;
Loh, Edmund ;
Gripenland, Jonas ;
Tiensuu, Teresa ;
Vaitkevicius, Karolis ;
Barthelemy, Mathieu ;
Vergassola, Massimo ;
Nahori, Marie-Anne ;
Soubigou, Guillaume ;
Regnault, Beatrice ;
Coppee, Jean-Yves ;
Lecuit, Marc ;
Johansson, Joergen ;
Cossart, Pascale .
NATURE, 2009, 459 (7249) :950-956
[24]   Specific and rapid detection of foodborne Salmonella by loop-mediated isothermal amplification method [J].
Wang, Li ;
Shi, Lei ;
Alam, M. J. ;
Geng, Yuhuan ;
Li, Lin .
FOOD RESEARCH INTERNATIONAL, 2008, 41 (01) :69-74
[25]   The Novel Multiple Inner Primers-Loop-Mediated Isothermal Amplification (MIP-LAMP) for Rapid Detection and Differentiation of Listeria monocytogenes [J].
Wang, Yi ;
Wang, Yan ;
Ma, Aijing ;
Li, Dongxun ;
Luo, Lijuan ;
Liu, Dongxin ;
Hu, Shoukui ;
Jin, Dong ;
Liu, Kai ;
Ye, Changyun .
MOLECULES, 2015, 20 (12) :21515-21531
[26]   Food safety incidents in the red meat industry: A review of foodborne disease outbreaks linked to the consumption of red meat and its products, 1991 to 2021 [J].
Warmate, Dein ;
Onarinde, Bukola A. .
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY, 2023, 398
[27]   Loop-mediated isothermal amplification (LAMP): a versatile technique for detection of micro-organisms [J].
Wong, Y. -P. ;
Othman, S. ;
Lau, Y. -L. ;
Radu, S. ;
Chee, H. -Y. .
JOURNAL OF APPLIED MICROBIOLOGY, 2018, 124 (03) :626-643
[28]   Loop-Mediated Isothermal Amplification for Salmonella Detection in Food and Feed: Current Applications and Future Directions [J].
Yang, Qianru ;
Domesle, Kelly J. ;
Ge, Beilei .
FOODBORNE PATHOGENS AND DISEASE, 2018, 15 (06) :309-331
[29]  
Zhao X, 2014, J MICROBIOL BIOTECHN, V24, P297