A Label-Free Fluorescent AND Logic Gate Aptasensor for Carbohydrate Antigen 15-3 Detection Based on Graphene Oxide

Author(s):  
Wenxiao Hu ◽  
Yafei Dong ◽  
Luhui Wang ◽  
Yue Wang ◽  
Mengyao Qian ◽  
...  

Background: Molecular logic gate always used fluorescent dyes to realize fluorescence signal. The labeling of the fluorophore is relatively expensive, low yield and singly labeled impuritiesaffects the affinity between the target and the aptamer. Label-free fluorescent aptamer biosensor strategy has attracted widespread interest due to lower cost and simple. Objective: Herein, we have designed a AND logic gate fluorescent aptasensor for detecting carbohydrate antigen 15-3(CA15-3) based on label-free fluorescence signal output. Materials and Methods: A hairpin DNA probe consists of CA15-3 aptamer and partly anti-CA15-3 aptamer sequences as a long stem and G-rich sequences of the middle ring as a quadruplex-forming oligomer. G-rich sequences can fold into a quadruplex by K+, and then G-quadruplex interacts specifically with N-methylmesoporphyrin IX(NMM), leading to a dramatic increase in fluorescence of NMM. With CA15-3 and NMM as the two inputs, the fluorescence intensity of the NMM is the output signal. Lacking of CA15-3 or NMM, there is no significant fluorescence enhancing, and the output of the signal is “0”. The fluorescence signal was dramatically increasing and the output of the signal is “1” only when CA15-3 protein and NMM were added at the same time. Results: This biosensor strategy possessed selectivity, high sensitivity for detecting CA15-3 protein from 10 to 500 U mL-1 and the detection limit was 10 U mL-1, and also showed good reproducibility in spiked human serum. Conclusion: In summary, the proposed AND logic gate fluorescent aptasensor could specifically detect CA15-3.

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jingjing Zhang ◽  
Handan Xu ◽  
Chunhui Li ◽  
Yilin Wang ◽  
Debing Liu ◽  
...  

Abstract Background In this paper, a simple, enzyme-free, label-free fluorescence, high sensitivity logic gate hairpin aptasensor was developed for adenosine triphosphate (ATP) detection based on graphene oxide (GO) and PicoGreen dye. Methods Using single-strand deoxyribonucleic acid (DNA) and adenosine triphosphate (ATP) as input signal and fluorescence signal as output signal, if single-strand DNA (DNA-L), single-strand DNA (DNA-S), and ATP were present at the same time, one segment of DNA-L formed a hairpin ring with ATP, and the other segment of DNA-L formed a completely complementary hairpin stem with DNA-S. The hairpin DNA was detached from the GO surface, and PicoGreen dye was embedded into the hairpin stem, and the fluorescence signal was enhanced. The molecular logic gate was constructed through the establishment of logic histogram, logic circuit, truth table, and logic formula. The biosensor-related performances including sensitivity, selectivity, and linearity were investigated, respectively. Results We have successfully constructed a AND logic gate. The detection limit of ATP is 138.0 pmol/L (3σ/slope) with detection range of 50–500 nmol/L (R2 = 0.98951), and its sensitivity is 4.748 × 106–6.875 × 108 a.u. (mol/L)−1. Conclusions The logic gate hairpin aptamer sensor has the advantages of high sensitivity, low detection limit, and low cost, and can be successfully applied to the detection of adenosine triphosphate (ATP) in actual human urine samples.


2011 ◽  
Vol 239-242 ◽  
pp. 934-939
Author(s):  
Hui Xu ◽  
Shuli Gao ◽  
Jian Nong Chen ◽  
Quan Wen Liu

We report a label-free, fast, fluorescence turn on assay for Hg2+detecton by using mercury-specific DNA (MSD), Sybr Green I (SG) and gold nanoparticles (AuNPs). SG efficiently discriminates MSD and MSD/Hg2+complex. The addition of gold nanoparticle decreases the background fluorescence signal further for MSD. The fluorescence intensity of MSD/Hg2+complex keeps constant after addition of AuNPs. This property improves the signal-to-background ratio and decreases the detection limitation further. In addition, the method shows improved selectivity compared with that in the absence of AuNPs. This strategy could be applied to the detection of potassium ions and showed good generality.


2021 ◽  
Author(s):  
Jun Yao ◽  
Li Wang

(A) Schematic diagram of the interaction and dual detection of Cu2+ and MPG by means of fluorescence “on–off–on” switching. (B) Molecular logic gate and truth table constructed based on Cu2+ and MPG as inputs and emission signal as output.


Sensors ◽  
2018 ◽  
Vol 18 (8) ◽  
pp. 2441 ◽  
Author(s):  
Xinxing Tang ◽  
Kefeng Wu ◽  
Han Zhao ◽  
Mingjian Chen ◽  
Changbei Ma

Adenosine deaminase (ADA), able to catalyze the irreversible deamination of adenosine into inosine, can be found in almost all tissues and plays an important role in several diseases. In this work, we developed a label-free fluorescence method for the detection of adenosine deaminase activity and inhibition. In the presence of ADA, ATP has been shown to be hydrolyzed. The ATP aptamer was shown to form a G-quadruplex/thioflavin T (ThT) complex with ThT and exhibited an obvious fluorescence signal. However, the ATP aptamer could bind with ATP and exhibited a low fluorescence signal because of the absence of ADA. This assay showed high sensitivity to ADA with a detection limit of 1 U/L based on an SNR of 3 and got a good linear relationship within the range of 1–100 U/L with R2 = 0.9909. The LOD is lower than ADA cutoff value (4 U/L) in the clinical requirement and more sensitive than most of the reported methods. This technique exhibited high selectivity for ADA against hoGG I, UDG, RNase H and λexo. Moreover, this strategy was successfully applied for assaying the inhibition of ADA using erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) and, as such, demonstrated great potential for the future use in the diagnosis of ADA-relevant diseases, particularly in advanced drug development.


2017 ◽  
Vol 5 (24) ◽  
pp. 4607-4613 ◽  
Author(s):  
Ting Hou ◽  
Tongxing Zhao ◽  
Wei Li ◽  
Feng Li ◽  
Panpan Gai

We proposed a label-free visual platform based on Cu–GMP CPNs for both self-correcting molecular logic gate construction and sensitive biosensing.


2004 ◽  
Vol 9 (6) ◽  
pp. 481-490 ◽  
Author(s):  
Brian T. Cunningham ◽  
Peter Li ◽  
Stephen Schulz ◽  
Bo Lin ◽  
Cheryl Baird ◽  
...  

Screening of biochemical interactions becomes simpler, less expensive, and more accurate when labels, such as fluorescent dyes, radioactive markers, and colorimetric reactions, are not required to quantify detected material. SRU Biosystems has developed a biosensor technology that is manufactured on continuous sheets of plastic film and incorporated into standard microplates and microarray slides to enable label-free assays to be performed with high throughput, high sensitivity, and low cost per assay. The biosensor incorporates a narrow band guided-mode resonance reflectance filter, in which the reflected color is modulated by the attachment/detachment of biochemical material to the surface. The technology offers 4 orders of linear dynamic range and uniformity within a plate, with a coefficient of variation of 2.5%. Using conventional biochemical immobilization surface chemistries, a wide range of assay applications are enabled. Small molecule screening, cell proliferation/cytotoxicity, enzyme activity screening, protein-protein interaction, and cell membrane receptor expression are among the applications demonstrated.


2019 ◽  
Author(s):  
Adam Eördögh ◽  
Carolina Paganini ◽  
Dorothea Pinotsi ◽  
Paolo Arosio ◽  
Pablo Rivera-Fuentes

<div>Photoactivatable dyes enable single-molecule imaging in biology. Despite progress in the development of new fluorophores and labeling strategies, many cellular compartments remain difficult to image beyond the limit of diffraction in living cells. For example, lipid droplets, which are organelles that contain mostly neutral lipids, have eluded single-molecule imaging. To visualize these challenging subcellular targets, it is necessary to develop new fluorescent molecular devices beyond simple on/off switches. Here, we report a fluorogenic molecular logic gate that can be used to image single molecules associated with lipid droplets with excellent specificity. This probe requires the subsequent action of light, a lipophilic environment and a competent nucleophile to produce a fluorescent product. The combination of these requirements results in a probe that can be used to image the boundary of lipid droplets in three dimensions with resolutions beyond the limit of diffraction. Moreover, this probe enables single-molecule tracking of lipids within and between droplets in living cells.</div>


The Analyst ◽  
2021 ◽  
Author(s):  
Ruirui Zhao ◽  
Lu Zhao ◽  
Haidi Feng ◽  
Xiaoliang Chen ◽  
Huilin Zhang ◽  
...  

Fluorescence sensing platforms based on HCR and G-quadruplex DNAzyme amplification strategies for the detection of prostate-specific antigen.


Author(s):  
Antonia Perju ◽  
Nongnoot Wongkaew

AbstractLateral flow assays (LFAs) are the best-performing and best-known point-of-care tests worldwide. Over the last decade, they have experienced an increasing interest by researchers towards improving their analytical performance while maintaining their robust assay platform. Commercially, visual and optical detection strategies dominate, but it is especially the research on integrating electrochemical (EC) approaches that may have a chance to significantly improve an LFA’s performance that is needed in order to detect analytes reliably at lower concentrations than currently possible. In fact, EC-LFAs offer advantages in terms of quantitative determination, low-cost, high sensitivity, and even simple, label-free strategies. Here, the various configurations of EC-LFAs published are summarized and critically evaluated. In short, most of them rely on applying conventional transducers, e.g., screen-printed electrode, to ensure reliability of the assay, and additional advances are afforded by the beneficial features of nanomaterials. It is predicted that these will be further implemented in EC-LFAs as high-performance transducers. Considering the low cost of point-of-care devices, it becomes even more important to also identify strategies that efficiently integrate nanomaterials into EC-LFAs in a high-throughput manner while maintaining their favorable analytical performance.


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