A real-time loop-mediated isothermal amplification assay for rapid detection of Shigella species
Tropical Biomedicine
; : 709-720, 2014.
Article
em En
| WPRIM
| ID: wpr-630429
Biblioteca responsável:
WPRO
ABSTRACT
Shigellosis is a foodborne illness caused by the genus Shigella and is an important global health issue. The development of effective techniques for rapid detection of this pathogen is essential for breaking the chain of transmission. Therefore, we have developed a novel loop-mediated isothermal amplification (LAMP) assay targeting the invasion plasmid antigen H (ipaH) gene to rapidly detect Shigella species. This assay could be performed in 90 min at an optimal temperature of 64ºC, with endpoint results visualized directly. Notably, the method was found to be more sensitive than conventional PCR. Indeed, the detection limit for the LAMP assay on pure bacterial cultures was 5.9 x 105 CFU/ml, while PCR displayed a limit of 5.9 x 107 CFU/ml. In spiked lettuce samples, the sensitivity of the LAMP assay was 3.6 x 104 CFU/g, whereas PCR was 3.6 x 105 CFU/g. Overall, the assay accurately identified 32 Shigella spp. with one enteroinvasive Escherichia coli displaying positive reaction while the remaining 32 non-Shigella strains tested were negative.
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WPRIM
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Revista:
Tropical Biomedicine
Ano de publicação:
2014
Tipo de documento:
Article