Robust tag-free aptasensor for monitoring of tobramycin: Architecting of rolling circle amplification and fluorescence synergism

Zahra Khoshbin; Noor Mohammad Danesh; Morteza Alinezhad Nameghi; Mohammad Ramezani; Mona Alibolandi; Mersedeh Shayan; Ali Samie; Khalil Abnous; Seyed Mohammad Taghdisi

doi:10.1016/j.ab.2023.115197

Robust tag-free aptasensor for monitoring of tobramycin: Architecting of rolling circle amplification and fluorescence synergism

Anal Biochem. 2023 Aug 1:674:115197. doi: 10.1016/j.ab.2023.115197. Epub 2023 May 30.

Authors

Zahra Khoshbin¹, Noor Mohammad Danesh², Morteza Alinezhad Nameghi², Mohammad Ramezani³, Mona Alibolandi³, Mersedeh Shayan², Ali Samie⁴, Khalil Abnous⁵, Seyed Mohammad Taghdisi⁶

Affiliations

¹ Targeted Drug Delivery Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Medicinal Chemistry, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
² Department of Medicinal Chemistry, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
³ Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran.
⁴ Targeted Drug Delivery Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Medicinal Chemistry, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
⁵ Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Medicinal Chemistry, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address: [email protected].
⁶ Targeted Drug Delivery Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address: [email protected].

PMID: 37263339
DOI: 10.1016/j.ab.2023.115197

Abstract

With the unpredictable risks on human health and ecological safety, tobramycin (TOB) as an extensively applied antibiotic has embraced global concern. Herein, a label-free fluorescent aptasensor was developed that opened up an innovative sensing strategy for monitoring trace TOB levels. Based on the rolling circle amplification (RCA) process, a giant DNA building was established by the catalytic action of T4 DNA ligase and Phi 29 DNA polymerase with the cooperation of the specific aptamer as a primer skeleton. By having the role of signal amplifier template, the RCA product with the G-quadruplex sequence duplications was decorated by a high number of the thioflavin T (ThT) fluorescent dyes. The aptasensor with good selectivity toward TOB achieved a detection limit as low as 150 pM. Thanks to its accurate target quantification, ease of operation, economic manufacture, as well as high potency for real-time and point-of-care testing, the represented aptasensor is superb for clinical application and food safety control.

Keywords: Aptamer; Biosensor; Fluorescence approach; Rolling circle amplification; Thioflavin T; Tobramycin.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aptamers, Nucleotide* / genetics
Biosensing Techniques*
DNA / genetics
DNA-Directed DNA Polymerase
Fluorescent Dyes
Humans
Limit of Detection
Nucleic Acid Amplification Techniques
Tobramycin

Substances

Tobramycin
DNA
DNA-Directed DNA Polymerase
Fluorescent Dyes
Aptamers, Nucleotide