Detecting and Discriminating Shigella sonnei Using an Aptamer-Based Fluorescent Biosensor Platform

Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry
Myeong-Sub SongYang-Hoon Kim

Abstract

In this paper, a Whole-Bacteria SELEX (WB-SELEX) strategy was adopted to isolate specific aptamers against Shigella sonnei. Real-time PCR amplification and post-SELEX experiment revealed that the selected aptmers possessed a high binding affinity and specificity for S. sonnei. Of the 21 aptamers tested, the C(t) values of the SS-3 and SS-4 aptamers (Ct = 13.89 and Ct = 12.23, respectively) had the lowest value compared to other aptamer candidates. The SS-3 and SS-4 aptamers also displayed a binding affinity (KD) of 39.32 ± 5.02 nM and 15.89 ± 1.77 nM, respectively. An aptamer-based fluorescent biosensor assay was designed to detect and discriminate S. sonnei cells using a sandwich complex pair of SS-3 and SS-4. The detection of S. sonnei by the aptamer based fluorescent biosensor platform consisted of three elements: (1) 5'amine-SS-4 modification in a 96-well type microtiter plate surface (N-oxysuccinimide, NOS) as capture probes; (2) the incubation with S. sonnei and test microbes in functionalized 96 assay wells in parallel; (3) the readout of fluorescent activity using a Cy5-labeled SS-3 aptamer as the detector. Our platform showed a significant ability to detect and discriminate S. sonnei from other enteric species such as ...Continue Reading

References

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May 23, 2017·Nanoscale·Simranjeet Singh SekhonYang-Hoon Kim

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Citations

Oct 14, 2017·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Harri Lönnberg
Feb 2, 2018·Advanced Materials·Yuezhou ZhangHongbo Zhang
Jul 13, 2019·Critical Reviews in Food Science and Nutrition·Wei WuQingli Yang
Jun 27, 2020·Applied Microbiology and Biotechnology·Fernanda Raquel Wust SchmitzDachamir Hotza
Jul 28, 2020·International Journal of Molecular Sciences·Nevina E Trunzo, Ka Lok Hong
Oct 14, 2020·Applied Microbiology and Biotechnology·Jiecan YiTianhan Kai
Oct 11, 2020·International Journal of Molecular Sciences·Chioma Izzi-EngbeayaWaljit S Dhillo
Jan 29, 2021·Analytical and Bioanalytical Chemistry·Lin ZhuChaoyong Yang
Mar 9, 2021·Frontiers in Microbiology·Higor Sette PereiraTiago Antônio de Oliveira Mendes
Mar 9, 2021·Frontiers in Microbiology·Mariam Saad, Sebastien P Faucher
Mar 20, 2018·ACS Combinatorial Science·Woo-Ri ShinYang-Hoon Kim
Jul 17, 2018·ACS Combinatorial Science·Se Hee LeeJi-Young Ahn

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Methods Mentioned

BETA
PCR
biosensor
Surface Plasmon
exponential enrichment
SELEX
chip
ELISA
Assay

Software Mentioned

ClustalX
BIA
MJ opticon monitor
Mfold

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