This study aimed to establish a method using reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) for rapid detection of Shigella sp.. Multiple sets of primers were designed according to the specific conserved gene ipaH of genus Shigella. An optimized reaction system was established, and its accuracy, specificity, as well as sensitivity were evaluated using artificial contaminated samples. The results showed that RT-LAMP assay successfully detected ipaH within 30 min under isothermal conditions at 65 °C. In the viable/damage bacteria model, this method even showed suitable accuracy better than that of the national standard method. Only four strains from Shigella sp. were specifically detected from a mixture of 23 different bacterial standard strains. The detection sensitivity of Shigella RNA was 7 fg/μL. The detection sensitivity of artificially contaminated skimmed milk samples reached 40 CFU/g, which was two orders of magnitude higher than the RT-PCR method. This RT-LAMP assay could accurately detect genus Shigella. This assay was rapid, specific and sensitive, which could be used for rapid screening and on-site monitoring of Shigella sp..
DING Mengxuan
,
LIU Xiu
,
LIU Boyang
,
LIANG Yulin
,
ZHOU Zhensen
,
YIN Jianjun
,
SONG Quanhou
. Rapid detection of Shigella sp. by reverse transcription loop-mediated isothermal amplification[J]. Food and Fermentation Industries, 2019
, 45(4)
: 193
-199
.
DOI: 10.13995/j.cnki.11-1802/ts.018815
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