An aptamer-array-based sample-to-answer biosensor for ochratoxin a detection via fluorescence resonance energy transfer

Yongning Li, Zhenfei Peng, Yaxi Li, Min Xiao, Gongjun Tan, Wenlian Wang, Yu Wang, Min Fang, Shu Zhang, Chenling Tang, Bowen Yang, Tianfu Wu

Research output: Contribution to journalArticlepeer-review

Abstract

Food toxins are a hidden threat that can cause cancer and tremendously impact human health. Therefore, the detection of food toxins in a timely manner with high sensitivity is of paramount importance for public health and food safety. However, the current detection methods are relatively time-consuming and not practical for field tests. In the present work, we developed a novel aptamer-chip-based sample-to-answer biosensor (ACSB) for ochratoxin A (OTA) detection via fluorescence resonance energy transfer (FRET). In this system, a cyanine 3 (Cy3)-labeled OTA-specific biotinylated aptamer was immobilized on an epoxy-coated chip via streptavidin-biotin binding. A complementary DNA strand to OTA aptamer at the 3-end was labeled with a black hole quencher 2 (BHQ2) to quench Cy3 fluorescence when in proximity. In the presence of OTA, the Cy3-labeled OTA aptamer bound specifically to OTA and led to the physical separation of Cy3 and BHQ2, which resulted in an increase of fluorescence signal. The limit of detection (LOD) of this ACSB for OTA was 0.005 ng/mL with a linearity range of 0.01–10 ng/mL. The cross-reactivity of ACSB against other mycotoxins, ochratoxin B (OTB), aflatoxin B1 (AFB1), zearalenone (ZEA), or deoxynilvalenol (DON), was less than 0.01%. In addition, this system could accurately detect OTA in rice samples spiked with OTA, and the mean recovery rate of the spiked-in OTA reached 91%, with a coefficient of variation (CV) of 8.57–9.89%. Collectively, the ACSB may represent a rapid, accurate, and easy-to-use platform for OTA detection with high sensitivity and specificity.

Original languageEnglish (US)
Article number309
JournalChemosensors
Volume9
Issue number11
DOIs
StatePublished - Nov 2021

Keywords

  • Aptamer
  • FRET
  • Ochratoxin A
  • On-chip assay

ASJC Scopus subject areas

  • Analytical Chemistry
  • Physical and Theoretical Chemistry

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