한빛사논문
- 조회855
Thi Thanh-Qui Nguyen, Eun Ryung Kim, Man Bock Gu*
Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul, Republic of Korea
*Corresponding author.
Abstract
A pair of aptamers for Staphylococcus aureus (S. aureus) is immensely needed for developing sandwich-type signal-on electrochemical aptasensors. In this study, we have successfully developed a cognate pair of aptamers that bind to S. aureus simultaneously, among many aptamer candidates screened out after a total of ten rounds of bacterial cell-based systemic evolution of ligands by exponential enrichment (SELEX). The obtained aptamer candidates have been estimated by using flow cytometry and confocal microscope, to evaluate their binding affinity and specificity to the target cells. The screening for sandwich-type binding of cognate pair of aptamers with S. aureus was conducted by enzyme-based colorimetric assay and confirmed by circular dichroism (CD), two-color fluorescence imaging analysis, additionally. The cognate pair of two aptamers, named SA37 and SA81, showed very good affinity and specificity to S. aureus with their dissociation constants (Kd) of 16.5 ± 3.4 nM and 14.47 ± 8.18 nM, respectively. These newly discovered cognate pair of aptamers have been very successfully implemented to develop a sandwich-type signal-on electrochemical biosensor with the limit of detection (LOD) of 39 CFUs and 414 CFUs in buffer and spiked tap water samples, respectively. This study showed that this cognate pair of aptamers-based detection of S. aureus enables simple, rapid, and robust biosensors for food safety management.
Keywords : Staphylococcus aureus, Bacterial cell-based SELEX, Cognate pair of aptamers, Electrochemical biosensor, Chronoamperometry
논문정보
- Research article
- 2022년 02월 (BRIC 등록일 2022-01-28)
- 국내 연구진
- 바이오·의료융합 > 바이오센싱 및 나노바이오물질
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