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

Song, Myeong-Sub; Sekhon, Simranjeet Singh; Shin, Woo-Ri; Kim, Hyung Cheol; Min, Jiho; Ahn, Ji-Young; Kim, Yang-Hoon

doi:10.3390/molecules22050825

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

Myeong-Sub Song

^1,†,

Simranjeet Singh Sekhon

^1,†,

^1,†,

²,

³,

^1,* and

^1,*

School of Biological Sciences, Chungbuk National University, 1 Chungdae-Ro, Seowon-Gu, Cheongju 28644, Korea

Technology Transfer Center, Korea Research Institute of Bioscience & Biotechnology, 125 Gwahak-Ro, Yuseong-Gu, Daejeon 34141, Korea

Department of Bioprocess Engineering, Chonbuk National University, 567 Baekje-daero, Deokjin-Gu Jeonju, Jeonbuk 54896, Korea

Authors to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Academic Editor: Harri Lönnberg

Molecules 2017, 22(5), 825; https://doi.org/10.3390/molecules22050825

Received: 19 April 2017 / Revised: 11 May 2017 / Accepted: 14 May 2017 / Published: 17 May 2017

(This article belongs to the Special Issue Synthesis and Applications of Oligonucleotide Conjugates)

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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 (K_D) 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 E. coli, Salmonella typhimurium and other Shigella species (S. flexneri, S. boydii). In this study, we demonstrated the feasibility of an aptamer sensor platform to detect S. sonnei in a variety of foods and pave the way for its use in diagnosing shigellosis through multiple, portable designs. View Full-Text

Keywords: shigellosis; aptamer-based fluorescent biosensor; whole cell-SELEX; DNA aptamer

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MDPI and ACS Style

Song, M.-S.; Sekhon, S.S.; Shin, W.-R.; Kim, H.C.; Min, J.; Ahn, J.-Y.; Kim, Y.-H. Detecting and Discriminating Shigella sonnei Using an Aptamer-Based Fluorescent Biosensor Platform. Molecules 2017, 22, 825. https://doi.org/10.3390/molecules22050825

AMA Style

Song M-S, Sekhon SS, Shin W-R, Kim HC, Min J, Ahn J-Y, Kim Y-H. Detecting and Discriminating Shigella sonnei Using an Aptamer-Based Fluorescent Biosensor Platform. Molecules. 2017; 22(5):825. https://doi.org/10.3390/molecules22050825

Chicago/Turabian Style

Song, Myeong-Sub, Simranjeet Singh Sekhon, Woo-Ri Shin, Hyung Cheol Kim, Jiho Min, Ji-Young Ahn, and Yang-Hoon Kim. 2017. "Detecting and Discriminating Shigella sonnei Using an Aptamer-Based Fluorescent Biosensor Platform" Molecules 22, no. 5: 825. https://doi.org/10.3390/molecules22050825

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

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