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ARS Home » Northeast Area » Wyndmoor, Pennsylvania » Eastern Regional Research Center » Molecular Characterization of Foodborne Pathogens Research » Research » Publications at this Location » Publication #317554

Title: A comparison of in-house real-time LAMP assays with a commercial assay for the detection of pathogenic bacteria

Author
item WANG, DEGUO - Xuchang University
item WANG, YONGZHEN - Xuchang University
item XIAO, FUGANG - Xuchang University
item GUO, WEIYUN - Xuchang University
item ZHANG, YONGQUIG - Xuchang University
item WANG, AIPING - Zhengzhou University
item Liu, Yanhong

Submitted to: Molecules
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/20/2015
Publication Date: 5/25/2015
Citation: Wang, D., Wang, Y., Xiao, F., Guo, W., Zhang, Y., Wang, A., Liu, Y. 2015. A comparison of in-house real-time LAMP assays with a commercial assay for the detection of pathogenic bacteria. Molecules. 20(6):9487-9495.

Interpretive Summary: Bacterial pathogens cause a variety of serious illnesses in humans and animals, and rapid and sensitive methods to detect the pathogens in food and other types of samples are needed to prevent infections. A technique called the loop-mediated isothermal amplification (LAMP) method was evaluated to detect pathogens known as Listeria monocytogenes, Salmonella, Staphylococcus aureus, Streptococcus agalactiae, and Escherichia coli. Twelve LAMP-based methods designed in house were compared to the use of a commercially-available LAMP-based kit to detect these bacteria. Results generated by the commercial kit were superior to those of the in-house assays since the in-house assays gave some false positive results. Thus, the commercial LAMP kit was rapid and sensitive and can potentially be used as a diagnostic method using clinical samples and to detect the pathogens in food.

Technical Abstract: Molecular detection of bacterial pathogens based on LAMP methods is a faster and simpler approach than conventional culture methods. Although different LAMP-based methods for pathogenic bacterial detection are available, a systematic comparison of these different LAMP assays has not been performed. In this paper, we compared 12 in-house real-time LAMP assays with a commercialized kit (Isothermal Master Mix) for the detection of Listeria monocytogenes, Salmonella spp, Staphylococcus aureus, Escherichia coli O157, E. coli O26, E. coli O45, E. coli O103, E. coli O111, E. coli O121, E. coli O145 and Streptococcus agalactiae. False-positive results were observed in all 12 in-house real-time LAMP assays, while all the negative controls of Isothermal Master Mix remained negative after amplification. The detection limit of Isothermal Master Mix for Listeria monocytogenes, Salmonella spp, Staphylococcus aureus, Escherichia coli O157, E. coli O26, E. coli O45, E. coli O103, E. coli O111, E. coli O121 and Streptococcus agalactiae was 1 pg, whereas the sensitivity of the commercialized kit for E. coli O145 was 100 pg. In conclusion, the 12 in-house real-time LAMP assays were impractical to use, while the commercialized kit Isothermal Master Mix was useful for the detection of most bacterial pathogens.