Aptamer-Based Fluorometric Ochratoxin A Assay Based on Photoinduced Electron Transfer

Zhao, Han; Xiang, Xinying; Chen, Mingjian; Ma, Changbei

doi:10.3390/toxins11020065

Cited by 26 publications

(15 citation statements)

References 35 publications

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“…The results showed that the hybridization of the complementary strands destroyed the structure of the G-quadruplex ( Figure S4 ), resulting in the inhibition of fluorescence-quenching. A previous study depicted an aptasensor utilizing guanines as a quencher based on PIET [ 22 ]. However, the quencher is different from the current study.…”

Section: Resultsmentioning

confidence: 99%

Development of Fluorescent Aptasensors Based on G-Quadruplex Quenching Ability for Ochratoxin A and Potassium Ions Detection

Yang

Chu

Zeng³

et al. 2022

Biosensors

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G-quadruplexes have received significant attention in aptasensing due to their structural polymorphisms and unique binding properties. In this work, we exploited the fluorescence-quenching properties of G-quadruplex to develop a simple, fast, and sensitive platform for fluorescence detection of ochratoxin A (OTA) and potassium ions (K+) with a label-free fluorophore and quencher strategy. The quenching ability of G-quadruplex was confirmed during the recognition process after the formation of the G-quadruplex structure and the quenching of the labeled fluorescein fluorophore (FAM). The fluorescence-quenching mechanism was studied by introducing specific ligands of G-quadruplex to enhance the quenching effect, to show that this phenomenon is due to photo-induced electron transfer. The proposed fluorescence sensor based on G-quadruplex quenching showed excellent selectivity with a low detection limit of 0.19 nM and 0.24 µM for OTA and K+, respectively. Moreover, we demonstrated that our detection method enables accurate concentration determination of real samples with the prospect of practical application. Therefore, G-quadruplexes can be excellent candidates as quenchers, and the strategy implemented in the study can be extended to an aptasensor with G-quadruplex.

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Section: Resultsmentioning

confidence: 99%

Development of Fluorescent Aptasensors Based on G-Quadruplex Quenching Ability for Ochratoxin A and Potassium Ions Detection

Yang

Chu

Zeng³

et al. 2022

Biosensors

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“…Super G™ Blocking Buffer was purchased from GRACE BIO-LABS Inc. (Bend, OR, USA). OTA aptamer 5 -biotin-GATCGGGTGTGGGTGG CGTAAAGGGAGCATCGGACA-Cy3-3 and the complementary strand 5 -BHQ2-TGTCCG ATGC-3 [28] were synthesized in bio basic Inc. (Amherst, NY, USA). Streptavidin was diluted with PBS buffer, and then 500 picoliters of the sample was printed onto an epoxycoated slide with a microarray printing robot (sciFLEXARRAYER S3; Scienion) at 25 • C in an atmosphere of 60% humidity, as described previously [29].…”

Section: Chemicals Reagents and Instrumentsmentioning

confidence: 99%

An Aptamer-Array-Based Sample-to-Answer Biosensor for Ochratoxin A Detection via Fluorescence Resonance Energy Transfer

Peng²,

et al. 2021

Chemosensors

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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.

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“…Second, highly stable aptamers with prominent target diversity can be easily synthesized at low cost. Owing to these outstanding merits, several aptamer-based methods for OTA determination have been developed, such as electrochemistry- [24,25], fluorescence- [26,27], and colorimetry-based assays [28,29]. The first OTA aptamer synthesized by Cruz-Aguado for OTA detection was reported in 2008 and this aptamer has been widely used in the construction of many biosensors [30].…”

Section: Introductionmentioning

confidence: 99%

Amplified Fluorescent Aptasensor for Ochratoxin A Assay Based on Graphene Oxide and RecJf Exonuclease

Zhao

Xiong

Yan

et al. 2020

Toxins

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In this study, we developed an aptamer-based fluorescent sensing platform for the detection of ochratoxin A (OTA) based on RecJf exonuclease-assisted signal amplification and interaction between graphene oxide (GO) and the OTA aptamer (OTA-apt). After optimizing the experimental conditions, the present aptamer-based sensing system can exhibit excellent fluorescent response in the OTA assay, with a limit of detection of 0.07 ng/mL. In addition to signal amplification, this strategy is also highly specific for other interfering toxins. Furthermore, this aptasensor can be reliably used for assessing red wine samples spiked with different OTA concentrations (2.4, 6 and 20 ng/mL). The proposed assay plays an important role in the field of food safety and can be transformed for detecting other toxins by replacing the sequence that recognizes the aptamer.

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Aptamer-Based Fluorometric Ochratoxin A Assay Based on Photoinduced Electron Transfer

Cited by 26 publications

References 35 publications

Development of Fluorescent Aptasensors Based on G-Quadruplex Quenching Ability for Ochratoxin A and Potassium Ions Detection

Development of Fluorescent Aptasensors Based on G-Quadruplex Quenching Ability for Ochratoxin A and Potassium Ions Detection

An Aptamer-Array-Based Sample-to-Answer Biosensor for Ochratoxin A Detection via Fluorescence Resonance Energy Transfer

Amplified Fluorescent Aptasensor for Ochratoxin A Assay Based on Graphene Oxide and RecJf Exonuclease

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