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Molecules 2015, 20(6), 9487-9495; doi:10.3390/molecules20069487

A Comparison of In-House Real-Time LAMP Assays with a Commercial Assay for the Detection of Pathogenic Bacteria

1
Henan Postdoctoral Research Base, Food and Bioengineering College, Xuchang University, Xuchang 461000, Henan, China
2
Public Lab Center, Xuchang University, Xuchang 461000, Henan, China
3
Life Science College, Zhengzhou University, Zhengzhou 450000, Henan, China
4
Molecular Characterization of Foodborne Pathogens Research Unit, Eastern Regional Research Center, Agricultural Research Service, United States Department of Agriculture, Wyndmoor, PA 19038, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Derek J. McPhee
Received: 12 April 2015 / Accepted: 20 May 2015 / Published: 25 May 2015
(This article belongs to the Section Molecular Diversity)
View Full-Text   |   Download PDF [658 KB, uploaded 25 May 2015]

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. View Full-Text
Keywords: real-time loop-mediated isothermal amplification (LAMP); specificity; sensitivity; pathogenic bacteria real-time loop-mediated isothermal amplification (LAMP); specificity; sensitivity; pathogenic bacteria
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Wang, D.; Wang, Y.; Xiao, F.; Guo, W.; Zhang, Y.; Wang, A.; Liu, Y. A Comparison of In-House Real-Time LAMP Assays with a Commercial Assay for the Detection of Pathogenic Bacteria. Molecules 2015, 20, 9487-9495.

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