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

Molecules

Deguo Wang³  Yongzhen Wang¹  Fugang Xiao³  Weiyun Guo³  Yongqing Zhang³  Aiping Wang²  Yanhong Liu⁴ 

[1]Public Lab Center, Xuchang University, Xuchang 461000, Henan, China

[2] E-Mail:

[3]Life Science College, Zhengzhou University, Zhengzhou 450000, Henan, China

[4] E-Mail:

[5]Henan Postdoctoral Research Base, Food and Bioengineering College, Xuchang University, Xuchang 461000, Henan, China

[6] E-Mails:

[7]Molecular Characterization of Foodborne Pathogens Research Unit, Eastern Regional Research Center, Agricultural Research Service, United States Department of Agriculture, Wyndmoor, PA 19038, USA

关键词: real-time loop-mediated isothermal amplification (LAMP);    specificity;    sensitivity;    pathogenic bacteria;   

DOI  :  10.3390/molecules20069487

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.