Format

Send to

Choose Destination
Mol Cells. 2016 Nov 30;39(11):807-813. Epub 2016 Nov 18.

Cell-SELEX Based Identification of an RNA Aptamer for Escherichia coli and Its Use in Various Detection Formats.

Author information

1
Global Research Laboratory (GRL) for RNAi Medicine, Department of Chemistry, Sungkyunkwan University (SKKU), Suwon 16419, Korea.
2
Department of Bioengineering, Dongguk University, Seoul 04620, Korea.
3
Department of Biomedical Engineering and School of Mechanical Engineering, Inje University, Gimhae 50834, Korea.

Abstract

Escherichia coli are important indicator organisms, used routinely for the monitoring of water and food safety. For quick, sensitive and real-time detection of E. coli we developed a 2'F modified RNA aptamer Ec3, by Cell-SELEX. The 31 nucleotide truncated Ec3 demonstrated improved binding and low nano-molar affinity to E. coli. The aptamer developed by us out-performs the commercial antibody and aptamer used for E. coli detection. Ec3(31) aptamer based E. coli detection was done using three different detection formats and the assay sensitivities were determined. Conventional Ec3(31)-biotin-streptavidin magnetic separation could detect E. coli with a limit of detection of 1.3 × 106 CFU/ml. Although, optical analytic technique, biolayer interferometry, did not improve the sensitivity of detection for whole cells, a very significant improvement in the detection was seen with the E. coli cell lysate (5 × 104 CFU/ml). Finally we developed Electrochemical Impedance Spectroscopy (EIS) gap capacitance biosensor that has detection limits of 2 × 104 CFU/mL of E. coli cells, without any labeling and signal amplification techniques. We believe that our developed method can step towards more complex and real sample application.

KEYWORDS:

Escherichia coli; aptamer; biolayer Interferometry; biosensor; cell-SELEX; impedimetric sensor

PMID:
27871171
PMCID:
PMC5125936
DOI:
10.14348/molcells.2016.0167
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Publishing M2Community Icon for PubMed Central
Loading ...
Support Center