Project description:We have developed a novel molecular logic gate system based on the incorporation of aptamer-crosslinked hydrogels. Modified gold nanoparticles are used as the output signal, which is visible to the naked eye. This system is designed for AND and OR operations using two chemicals as stimulus inputs.
Project description:Excessive residue of metronidazole (MNZ) in food is harmful to the human body. There is an urgent demand to develop a portable tool for MNZ detection on-site. In this study, fifteen aptamers were prepared through targeted base mutation. Apt1-3 with the highest enrichment was chosen for further study. Its affinity was characterized by molecular docking simulation, AuNPs colorimetric assay, graphene oxide (GO) fluorescence assay, and exonuclease assay. Kd was determined by GO fluorescence assay (Kd: 92.60 ± 25.59 nM). Its specificity was also characterized by an exonuclease assay. A novel aptasensor was constructed by using the newly identified aptamer combined with the smartphone dark box. The principle of color change is caused by the aggregation state of AuNPs. Smartphones act as reading instruments. The detection can be completed in just a few seconds without the aid of instruments, achieving a detection limit of 0.15 nmol/mL and a range of 6.7-44.4 nmol/mL (R 2 = 0.9810). Therefore, the constructed smartphone colorimetric sensor based on mutant aptamers has important applications in food detection.
Project description:In this paper, we describe an aptamer-based colorimetric assay (ABCA), which integrates enzyme-loaded microparticles for signal amplification with distance measurement for equipment-free quantitative readout. The distance measurement readout is on the basis of target-induced selective reduction in viscosity of reaction solution. Its utility is well demonstrated with inexpensive, sensitive, and selective detection of adenosine (model analyte) in buffer samples and real samples of human serum and urine with the naked eye. This ABCA method just requires operators to simply count the number of colored distance-relevant marked bars on the calibrated glass microsyringes (testing containers) to provide quantitative results. It thus holds great promise for wide applications particularly in limited-resource settings.
Project description:BackgroundAn aptamer based biosensor (aptasensor) was developed and evaluated for rapid colorimetric detection of Escherichia coli (E. coli) O157:H7.Methodology/principal findingsThe aptasensor was assembled by modifying the truncated lipopolysaccharides (LPS)-binding aptamer on the surface of nanoscale polydiacetylene (PDA) vesicle using peptide bonding between the carboxyl group of the vesicle and the amine group of the aptamer. Molecular recognition between E. coli O157:H7 and aptamer at the interface of the vesicle lead to blue-red transition of PDA which was readily visible to the naked eyes and could be quantified by colorimetric responses (CR). Confocal laser scanning microscope (CLSM) and transmission electron microscopy (TEM) was used to confirm the specific interactions between the truncated aptamer and E. coli O157:H7. The aptasensor could detect cellular concentrations in a range of 10(4)~ 10(8) colony-forming units (CFU)/ml within 2 hours and its specificity was 100% for detection of E. coli O157:H7. Compared with the standard culture method, the correspondent rate was 98.5% for the detection of E. coli O157:H7 on 203 clinical fecal specimens with our aptasensor.ConclusionsThe new aptasensor represents a significant advancement in detection capabilities based on the combination of nucleic acid aptamer with PDA vesicle, and offers a specific and convenient screening method for the detection of pathogenic bacteria. This technic could also be applied in areas from clinical analysis to biological terrorism defense, especially in low-resource settings.
Project description:Residue and illegal addition of Dexamethasone (DEX) in food has received widespread attention over the past few decades. Long-term intake of DEX will have a strong endocrine-disrupting effect, and there is an urgent need to develop highly sensitive and rapid on-site detection methods. In this work, a colorimetric sensor based on an unmodified aptamer and gold nanoparticles (Au NPs) was designed to detect DEX in milk and glucosamine. Under optimized conditions, the absorbance ratio of Au NPs increased linearly with DEX concentration over the range of 10-350 nmol/mL (r2 = 0.997), with a limit of detection (LOD) of 0.5 nmol/mL, and the recoveries ranged from 93.6 to 117%. To explore the interaction mechanism between aptamer and DEX, molecular docking and molecular dynamics simulations were applied to probe intermolecular interactions and structures of the complex. The establishment of aptamer-based sensors effectively avoids the antibody screening response, with a cost-efficient, excellent selective and great potential in DEX determination.
Project description:A highly selective and sensitive method for Cd(II) detection was developed based on aptamer and gold nanoparticles (AuNPs) combined with a colorimetric smartphone readout. The experimental conditions such as reaction time of polydiene dimethyl ammonium chloride (PDDA) and AuNPs, PDDA dose, time of aptamer and PDDA incubation, and aptamer concentration were optimized. Under the optimized conditions, the color and red(R) value of the solution was concentration-dependent on Cd(II). The proposed method exhibited a linear range of 1-400 ng/mL (r2 = 0.9794) with a limit of detection (LOD) of 1 ng/mL. This method had been successfully applied to test and quantify Cd(II) in water and rice samples, and the results were in full agreement with those from the atomic absorption spectrometer. Therefore, low-cost colorimetry demonstrated its potential for practical application in visual or quantitative detection with a smartphone. This approach can be readily applied to other analytes.
Project description:Contamination of milk by mycotoxins is a serious problem worldwide. Herein, we focused on the detection of aflatoxin B1 (AflB1) using a paper microfluidic device fabricated with specific aptamers as biorecognition elements. The fabrication process resulted in the generation of a leak proof microfluidic device where two zones were designed: control and test. Detection is achieved by color change when aflatoxin reacts with an aptamer followed by salt induced aggregation of gold nanoparticles. Specific aptamers for aflatoxin B1 were immobilized successfully onto the surface of gold nanoparticles. Biophysical characterization of the conjugated AuNPs-aptamer was done by UV-vis spectroscopy, DLS (dynamic light scattering), TEM (transmission electron microscopy). Under optimal conditions, the microfluidic device showed an excellent response for aflatoxin B1 detection in the range of 1 pM to 1 μM with a detection limit of up to 10 nM in spiked samples. Disadvantages associated with conventional techniques of ELISA were overcome by this one step detection technique with low operation cost, simple instrumentation, and user-friendly format with no interference due to chromatographic separation. The developed microfluidic paper-based analytical device (μPAD) can provide a tool for on-site detection of food toxins in less than a minute which is the main requirement for both qualitative and quantitative analysis in food safety and environmental monitoring.
Project description:Here, we designed a simple, rapid, and ultrasensitive colorimetric aptasensor for detecting anatoxin-a (ATX-a). The sensor employs a DNA aptamer as the sensing element and gold nanoparticles (AuNPs) as probes. Adsorption of the aptamer onto the AuNP surface can protect AuNPs from aggregation in NaCl solution, thus maintaining their dispersion state. In the presence of ATX-a, the specific binding of the aptamer with ATX-a results in a conformational change in the aptamer, which facilitates AuNP aggregation and, consequently, a color change of AuNPs from red to blue in NaCl solution. This color variation is directly associated with ATX-a concentration and can be easily measured using a UV/Vis spectrophotometer. The absorbance variation is linearly proportional to ATX-a concentration across the concentration range of 10 pM to 200 nM, with a detection limit of 4.45 pM and high selectivity against other interferents. This strategy was successfully applied to the detection of ATX-a in lake water samples. Thus, the present aptasensor is a promising alternative method for the rapid detection of ATX-a in the environment.
Project description:Exosomes constitute an emerging biomarker for cancer diagnosis because they carry multiple proteins that reflect the origins of parent cells. Assessing exosome surface proteins provides a powerful means of identifying a combination of biomarkers for cancer diagnosis. We report a sensor platform that profiles exosome surface proteins in minutes by the naked eye. The sensor consists of a gold nanoparticle (AuNP) complexed with a panel of aptamers. The complexation of aptamers with AuNPs protects the nanoparticles from aggregating in a high-salt solution. In the presence of exosomes, the non-specific and weaker binding between aptamers and the AuNP is broken, and the specific and stronger binding between exosome surface protein and the aptamer displaces aptamers from the AuNP surface and results in AuNP aggregation. This aggregation results in a color change and generates patterns for the identification of multiple proteins on the exosome surface.
Project description:We report a simple naked eye colorimetric detection assay developed for the small molecule creatinine using the surface passivation of gold nanoparticles (AuNPs) which is conjugated with a creatinine binding aptamer. The selective binding of creatinine to aptamer sequences causes a decrease in the catalytic activity of AuNPs, and the color change time of the 4-nitrophenol reduction was used for the quantitative colorimetric detection of creatinine. Herein, the surfaces of AuNPs acted as the catalyst for the reduction of 4-nitrophenol (yellow) to 4-aminophenol (colorless), and the passivation with creatinine bound aptamer sequences delayed the reduction. The developed assay was able to detect creatinine in a linear range of 2-20 mM with a limit of detection of 0.87 mM. The developed colorimetric assay was very selective and repeatable and could detect creatinine in the presence of interfering biomolecules. Moreover, the assay showed excellent results for the analysis of creatinine in artificial urine samples. The developed assay can be used as a point of care (POC) device for the naked eye detection of creatinine within few minutes without any instrument support.