期刊
BIOSENSORS & BIOELECTRONICS
卷 198, 期 -, 页码 -出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2021.113835
关键词
Staphylococcus aureus; Bacterial cell-based SELEX; Cognate pair of aptamers; Electrochemical biosensor; Chronoamperometry
类别
资金
- Nano.Material Technology Development Program through the National Research Foundation of Korea (NRF) - Ministry of Science, ICT and Future Planning [2016M3A7B4910555]
- National Research Foundation of Korea (NRF) - Korea government (MSIT) [2021R1A2C2013396]
- National Research Foundation of Korea [2021R1A2C2013396, 2016M3A7B4910555] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
A cognate pair of aptamers for Staphylococcus aureus was successfully developed through SELEX technology and implemented in a sandwich-type signal-on electrochemical biosensor, showing good affinity and specificity for S. aureus detection in food safety management.
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.
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