A real-time loop-mediated isothermal amplification assay for rapid detection of Shigella species
- Author:
Liew, P.S.
;
Teh, C.S.J.
;
Lau, Y.L.
;
Thong, K.L.
- Publication Type:Journal Article
- From:Tropical Biomedicine
2014;31(4):709-720
- CountryMalaysia
- Language:English
-
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.
- Full text:W020150601574632808836.pdf
