scholarly journals Graphene Oxide and Fluorescent Aptamer Based Novel Biosensor for Detection of 25-hydroxyvitamin D3

2021 ◽  
Author(s):  
Ritika Gupta ◽  
Sunaina Kaul ◽  
Vishal Singh ◽  
Sandeep Kumar ◽  
Nitin Kumar Singhal
Keyword(s):  
Vitamin D ◽  
Graphene Oxide ◽  
Limit Of Detection ◽  
Low Cost ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Detection Methods ◽  
Hydroxyvitamin D ◽  
25 Hydroxyvitamin D ◽  
Sensitive Sensor

Abstract For maintaining the healthy metabolic status, vitamin D is a beneficial metabolite stored majorly in its pre-activated form, 25-hydroxyvitamin D3 (25(OH)D3). Due to its important role in bone strengthening, the study was planned to quantify 25(OH)D3 levels in our blood. Quantification techniques for 25(OH)D3 are costly thus requiring a need for a low cost, and sensitive detection methods. In this work, an economic, and sensitive sensor for the detection of 25(OH)D3 was developed using aptamer and graphene oxide (GO). Aptamer is an oligonucleotide, sensitive towards its target, whereas, GO with 2D nanosheets provides excellent quenching surface. Aptamer labeled with fluorescein (5’, 6-FAM) is adsorbed by π -π interaction on the GO sheets leading to quenching of the fluorescence due to Förster resonance energy transfer (FRET). However, in the presence of 25(OH)D3, a major portion of aptamer fluorescence remains unaltered, due to its association with 25(OH)D3. However, in the absence, aptamer fluorescence gets fully quenched. Fluorescence intensity quenching was monitored using fluorescence spectrophotometer and agarose gel based system. The limit of detection of 25(OH)D3 by this method was found to be 0.15 µg/mL. Therefore, this method could come up as a new sensing method in the field of vitamin D detection.

scholarly journals Graphene oxide and fluorescent aptamer based novel biosensor for detection of 25-hydroxyvitamin D3

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ritika Gupta ◽  
Sunaina Kaul ◽  
Vishal Singh ◽  
Sandeep Kumar ◽  
Nitin Kumar Singhal
Keyword(s):  
Vitamin D ◽  
Graphene Oxide ◽  
Limit Of Detection ◽  
Low Cost ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Detection Methods ◽  
Hydroxyvitamin D ◽  
25 Hydroxyvitamin D ◽  
Sensitive Sensor

AbstractFor maintaining the healthy metabolic status, vitamin D is a beneficial metabolite stored majorly in its pre-activated form, 25-hydroxyvitamin D3 (25(OH)D3). Due to its important role in bone strengthening, the study was planned to quantify 25(OH)D3 levels in our blood. Quantification techniques for 25(OH)D3 are costly thus requiring a need for a low cost, and sensitive detection methods. In this work, an economic, and sensitive sensor for the detection of 25(OH)D3 was developed using aptamer and graphene oxide (GO). Aptamer is an oligonucleotide, sensitive towards its target, whereas, GO with 2D nanosheets provides excellent quenching surface. Aptamer labeled with fluorescein (5’, 6-FAM) is adsorbed by π–π interaction on the GO sheets leading to quenching of the fluorescence due to Förster resonance energy transfer (FRET). However, in the presence of 25(OH)D3, a major portion of aptamer fluorescence remains unaltered, due to its association with 25(OH)D3. However, in the absence, aptamer fluorescence gets fully quenched. Fluorescence intensity quenching was monitored using fluorescence spectrophotometer and agarose gel based system. The limit of detection of 25(OH)D3 by this method was found to be 0.15 µg/mL whereas when GO-COOH was used, limit of detection was improved to 0.075 µg/mL. Therefore, this method could come up as a new sensing method in the field of vitamin D detection.

scholarly journals Characterization of an Aptamer Directed against 25-Hydroxyvitamin D for the Development of a Competitive Aptamer-Based Assay

Biosensors ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 134 ◽  
Author(s):  
Marc Prante ◽  
Torsten Schüling ◽  
Bernhard Roth ◽  
Kort Bremer ◽  
Johanna Walter
Keyword(s):  
High Performance ◽  
Limit Of Detection ◽  
Detection Methods ◽  
Microarray Experiments ◽  
Hydroxyvitamin D ◽  
Detection Approach ◽  
Biosensor System ◽  
25 Hydroxyvitamin D ◽  
Current Detection

Detection of the small molecule 25-hydroxyvitamin D (25(OH)D) as the most relevant marker for vitamin D supply suffers from a high variability of results using the current detection methods, such as high-performance liquid chromatography (HPLC) and immunoassays. A new detection approach using a highly specific aptamer directed against 25(OH)D was established in this study based on the target-induced dissociation (TID) sensing approach. In this work, the aptamer was investigated regarding its structural properties as well as its binding affinity by using microscale thermophoresis (MST). Moreover, complementary oligonucleotides were designed based on the aptamer structure and were evaluated in MST experiments. Binding experiments of immobilized aptamers were conducted in microarray experiments. It could be shown that the aptamer exhibited the usual B-DNA structure and did not form any G-quadruplexes. The design of complementary oligonucleotides for the TID assay identified a putative 25(OH)D binding site within the aptamer. The limit of detection of the established competitive assay was determined to be 5.4 nM, which sets the stage for the development of a biosensor system.

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

Chemosensors ◽  
2021 ◽  
Vol 9 (11) ◽  
pp. 309
Author(s):  
Yongning Li ◽  
Zhenfei Peng ◽  
Yaxi Li ◽  
Min Xiao ◽  
Gongjun Tan ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Ochratoxin A ◽  
Fluorescence Resonance Energy Transfer ◽  
Limit Of Detection ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
High Sensitivity ◽  
Detection Methods ◽  
Fluorescence Signal ◽  
Fluorescence Resonance

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.

scholarly journals A Microfluidic Aptamer-Based Sensor for Detection of Mercury (II) and Lead (II) Ions in Water

2021 ◽  
Author(s):  
Wei-Hao Huang ◽  
Ruo-Yin Wu ◽  
Ko-Li Yeh ◽  
Van-Phung Mai ◽  
Ruey-Jen Yang
Keyword(s):  
Heavy Metal ◽  
Drinking Water ◽  
Graphene Oxide ◽  
Energy Transfer ◽  
Fluorescent Dyes ◽  
Limit Of Detection ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
World Health ◽  
The World

Heavy metal contaminants have serious consequences for the environment and human health. Consequently, effective methods for detecting their presence, particularly in water and food, are urgently required. Accordingly, the present study proposes a sensor for the detection of mercury Hg(II) and lead Pb(II) ions using graphene oxide (GO) as a quenching agent and aptamer solu-tion as a reagent. In the proposed device, the aptamer sequences are labeled by FAM and HEX fluorescent dyes, respectively, and are mixed with 500 ppm GO solution in a microfluidic device. The presence of Hg(II) and Pb(II) ions is then detected by measuring the change in the fluores-cence intensity of the GO/aptamer suspension as the aptamer molecules undergo fluorescence resonance energy transfer (FRET). The experimental results show that the aptamer sensors have a linear range of 10~250 nM (i.e., 2.0~50 ppb) for Hg(II) ions and 10~100 nM (i.e., 2.1~20.7 ppb) for Pb(II) ions. Furthermore, the limit of detection is around 2 ppb for both metals, which is signifi-cantly lower than the maximum limits of 6 ppb and 10 ppb prescribed by the World Health Or-ganization (WHO) for Hg(II) and Pb(II) in drinking water, respectively.

scholarly journals A Simple Fluorescent Aptasensing Platform Based on Graphene Oxide for Dopamine Determination

2021 ◽  
Author(s):  
Ahlem Teniou ◽  
amina rhouati ◽  
Gaëlle Catanante
Keyword(s):  
Graphene Oxide ◽  
Limit Of Detection ◽  
Rapid Determination ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
High Sensitivity ◽  
Fluorescence Recovery ◽  
Serum Samples ◽  
Experimental Conditions ◽  
Minor Response

Abstract Dopamine (DA) is a catecholamine neurotransmitter playing an important role in different biological functions including central nervous, renal, cardiovascular, and hormonal systems. The sensitive and selective detection of this neurotransmitter plays a key role in the early diagnosis of various diseases related to abnormal levels of dopamine. Therefore, it is of great importance to explore rapid, simple, and accurate methods for detection of dopamine with high sensitivity and specificity. We propose in this work, a fluorescent aptasensor based on graphene oxide (GO) as a quencher, for the rapid determination of dopamine. The principle of this aptasensor is based on fluorescence resonance energy transfer (FRET), where GO was used as energy donor, and a carboxyfluorescein (FAM)-labeled aptamer as acceptor. In the absence of DA, FAM-aptamer was adsorbed on the surface of GO through π-π stacking interactions between nucleotide bases and the carbon network, leading to a weak FRET and a quenching of the FAM fluorescence. However, by adding the target, the aptamer undergoes a conformational change to bind to DA with high affinity, resulting in a fluorescence recovery. Under the optimal experimental conditions, the fluorescence recovery was linearly proportional to the concentration of DA in the range of 3-1680 nM, with a limit of detection of 0.031 nM. Moreover, the developed assay exhibited minor response in the presence of various interferents and it revealed a satisfactory applicability in human serum samples.

Depressed Serum 25-Hydroxyvitamin D Levels Increase Hospital Stay and Alter Glucose Homeostasis in First-ever Ischemic Stroke

2019 ◽  
Vol 16 (4) ◽  
pp. 340-347
Author(s):  
Yuge Wang ◽  
Yanqiang Wang ◽  
Bingjun Zhang ◽  
Yinyao Lin ◽  
Sha Tan ◽  
...  
Keyword(s):  
Vitamin D ◽  
Ischemic Stroke ◽  
Hospital Stay ◽  
Functional Outcome ◽  
Vitamin D Deficiency ◽  
Glucose Homeostasis ◽  
Clinical Severity ◽  
Hydroxyvitamin D ◽  
25 Hydroxyvitamin D ◽  
Deficiency Group

Background and Objective: Vitamin D deficiency is internationally recognized among the potentially modifiable risk factors for ischemic cardio-cerebrovascular diseases. However, the association between vitamin D deficiency and stroke morbidity or mortality remains insufficiently known. Our aim is to investigate their relevance to 25-hydroxyvitamin D [25(OH) D] levels and clinical severity and outcome after 3 months in first-ever ischemic stroke. Methods: Retrospective analysis of 356 consecutive patients in first-ever ischemic stroke between 2013 and 2015. Serum 25(OH) D levels were measured at baseline. Stroke severity was assessed at admission using the National Institutes of Health Stroke Scale (NIHSS) score. Functional outcome after 3 months of onset was evaluated using the modified Rankin scale (mRS). Results: Among the 356 enrolled patients, HbA1c was higher in insufficiency/deficiency group than that in the sufficiency group (6.3 ± 1.7 vs. 5.9 ± 1.1, p =0.015). The hospital stay was longer in insufficiency/deficiency group than that in the sufficiency group (11 (8-17) vs. 9.5 (7-13), p = 0.035). There was a significant inversed trend between serum 25(OH) D levels and hospital stay (OR 0.960, P = 0.031), using logistic regression. Conclusions: 25(OH)D levels are associated with glucose homeostasis, 25(OH) D contributes to increase the length of hospital stay. Low serum 25-OHD level is an independent predictor for hospital stay in first-ever ischemic stroke. Vitamin D deficiency did not predict functional outcome in the span of 3 months.

scholarly journals Role of glucuronidated 25-hydroxyvitamin D on colon gene expression in mice

2020 ◽  
Vol 319 (2) ◽  
pp. G253-G260
Author(s):  
Carmen J. Reynolds ◽  
Nicholas J. Koszewski ◽  
Ronald L. Horst ◽  
Donald C. Beitz ◽  
Jesse P. Goff
Keyword(s):  
Gene Expression ◽  
Vitamin D ◽  
Hydroxyvitamin D ◽  
25 Hydroxyvitamin D

We found that 25OHD-Gluc, an endogenously produced metabolite, is delivered to the colon via bile to induce vitamin D-mediated responses in the colon.

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