A rapid diagnostic method for E. coli serogroups responsible for gastro-intestinal diseases using loop-mediated isothermal amplification

• Published in: Analytical methods 2015-01, Vol.7 (1), p.287-295
• Main Authors: Carnevale, Matthew L, Roche, Philip J. R, Najih, Mohamed, Paliouras, Miltiadis, Beitel, Lenore K, Trifiro, Mark A
• Format: Article
• Language: English
• Subjects: Amplification; Diagnostic systems; Escherichia; Genes; Instrumentation; Lamps; Pathogens; Running
• ISSN: 1759-9660; 1759-9679
• DOI: 10.1039/c4ay02686a

A rapid diagnostic method for Escherichia coli serogroup identification was developed, employing the loop-mediated isothermal amplification reaction (LAMP). Identifying the serogroup responsible for infection is accomplished using primers that are specific to sequences that are exclusive to the particular serogroup. Previous work has primarily focused on detecting one specific sequence or gene as a method of detection of a pathogenic entity. The novel approach involves using primers specific to various genes to determine which are present as a method of identification of the unknown pathogen rather than simple detection. The experiments conducted involved running the LAMP reaction and detecting amplification using gel electrophoresis, fluorescence, and localized surface plasmon resonance. The results obtained demonstrate that the LAMP reaction is efficient, and specific in that it only amplifies target DNA, and that it requires minimal instrumentation in comparison to various other nucleic acid amplification methods. This assay's principle and instrumentation is suited to resource-limited environments and when mobility is required for testing. The LAMP reaction as opposed the traditionally used PCR reaction can be used for molecular diagnostics of bacterial infections. Fluorescence based visualization allows ease of use in developing countries.