- 标题
- 摘要
- 关键词
- 实验方案
- 产品
-
Enzyme-free fluorometric assay for chloramphenicol based on double stirring bar-assisted dual signal amplification
摘要： An enzyme-free fluorometric assay is described that accomplishes dual signal amplification by making use of a two stirring bars. Two Y-shaped DNA probes were designed and placed on the bars. When the target (with chloramphenicol as model analyte) is added, it triggers target recycling and simultaneously catalyzes hairpin assembly (CHA). A large fraction of DNA primers is released by the analyte from the bar to the supernatant and open hairpins with G-quadruplex DNA sequence. The G-quadruplex can specifically bind thioflavin T (ThT) to emit fluorescence (with excitation/emission maxima at 445 and 485 nm) for quantification of chloramphenicol. An enzyme is not needed. ThT is added to the system as a fluorescent DNA probe. All this strongly reduces the cost for sensor construction and usage. The dual signal amplification steps occur simultaneously which reduces the detection time. The assay was successfully employed to the determination of CAP in spiked milk and fish samples within 60 min and with a 16 pM limit of detection (at S/N = 3).
关键词： Food safety,Antibiotics detection,Thioflavin T,Catalyzed hairpin assembly,Target recycling
更新于2025-11-19 16:46:39
-
Zero background and triple-signal amplified fluorescence aptasensor for antibiotics detection in foods
摘要： It's important to eliminate matrix interference for accurate detecting antibiotic residues in complex food samples. In this study, we designed a zero-backgrounded fluorescence aptasensor to achieve on-site detection of antibiotic residues, with chloramphenicol (CAP) as representative analyte. Moreover, a three stir-bars assisted target recycling system (TSBTR) was designed to achieve triple signal amplification and increase the sensitivity. The bars included one magnetic stir-bar modified with two kinds of long DNA chains, and two gold stir-bars modified with Y shape-duplex DNA probes respectively. In the presence of CAP, the target could recurrently react with the probes on the bars and replace a large amount of long DNA chains into supernatant. After then, the bars were taken out and SYBR green dye was added to the solution. The dye can specifically intercalate into the duplex structures of DNA chains to emit fluorescence while not emitting a signal in its free state. Under the optimized experimental conditions, a wide linear response range of 5 orders of magnitude from 0.001 ng mL?1 to 10 ng mL?1 was achieved with a detection limit of 0.033 pg mL?1 CAP. The assay was successfully employed to detect CAP in food samples (milk & fish) with consistent results with ELISA's. High selectivity and sensitivity were attributed to the zero background signal and triple signal-amplification strategy. Moreover, the detection time can be shortened to 40 min due to that three signal amplified process can occur simultaneously. The fluorescent aptasensor was also label- and enzyme-free. All these ensure the platform to be rapid, cost-effective, easily-used, and is especially appropriate for detection antibiotics in food safety.
关键词： Three stir-bars assisted target recycling,Triple signal amplification,Zero background signal,Fluorescence aptasensor,Antibiotics
更新于2025-11-19 16:46:39
-
Building a Fluorescent Aptasensor Based on Exonuclease-Assisted Target Recycling Strategy for One-Step Detection of T-2 Toxin
摘要： In this work, a rapid and accurate assay was successfully developed for T-2 toxin detection based on exonuclease-catalyzed target recycling strategy. Upconversion nanoparticles (UCNPs) were conjugated with T-2 aptamer and used as signal probes, while magnetic nanoparticles (MNPs) were conjugated with the complementary DNA of T-2 aptamer (cDNA) and used as capture probes. The results reveled that good linear correlation (R2 = 0.9988) was achieved for T-2 toxin detection over the concentration range of 0.1–100 ng/mL with a detection limit as low as 0.035 ng/mL (S/N = 3). In addition, the reliability of the proposed method was also applied to the determination of T-2 toxin contents in real food samples and the average recoveries ranged from 95.97 to 104.00%. The sensing platform developed in our study demonstrated great potential for simple and sensitive detection of T-2 toxin contents in food samples.
关键词： T-2 toxin,Food safety,Fluorescence,Aptasensor,Target recycling strategy
更新于2025-09-23 15:23:52
-
Ultrasensitive and label-free detection of ATP by using gold nanorods coupled with enzyme assisted target recycling amplification
摘要： Abnormal concentration of adenosine triphosphate (ATP) is directly asscociate with several diseases. Thus, sensitive detection of ATP is essential to early diagnosis of disease. Herein, we described an ultrasensitive strategy for ATP detection by using positively charged gold nanorods ((+)AuNPs) as an efficient fluorescence quenching platform, coupled with exonuclease (cid:1) (Exo (cid:1)) assisted target recycling amplification. To construct the sensor, DNA template that contained ATP aptamer was used for the formation of AgNCs signal probe (DNA/AgNCs), the structure of it could change to duplex after the interaction of it with ATP. Such DNA template or duplex DNA product could electrostatically adsorb onto (+)AuNRs surface, resulting in the quenching of the fluorescence signal due to the vicinity of AgNCs to (+)AuNRs. With the addition of Exo (cid:1), DNA duplex could be hydrolyzed and released from (+)AuNRs surface, leading to the recovery of a strong fluorescent signal, while ATP could be regenerated for next target recycling. Combing the good fluorescence quenching ability of (+)AuNRs and the Exo (cid:1) assisted signal amplification, a low detection limit of 26 pM was achieved for ATP detection. Notably, the proposed method can be successfully applied for detecting ATP in serum samples, indicating a potential application value in early cancer diagnosis.
关键词： Exo (cid:1),Fluorescent sensor,(+)AuNRs,DNA/AgNCs,ATP detection,Target recycling amplification
更新于2025-09-16 10:30:52
-
An enzyme-free FRET nanoprobe for ultrasensitive ketamine detection based on ATP-fueled target recycling
摘要： Ketamine is a commonly abused drug due to its stimulant, dissociative and hallucinogenic effects. An overdose of ketamine has been found to cause a variety of side effects. Therefore, the identification and quantification of ketamine are of significant importance for clinical purposes and drug seizing. However, conventional methods for ketamine detection possess some disadvantages such as sophisticated procedures, expensive instruments and low sensitivity. Herein, we develop a novel fluorescent nanoprobe for ultrasensitive ketamine detection with signal amplification based on Adenosine Triphosphate (ATP)-fueled target recycling and FRET (fluorescence resonance energy transfer) occurring between the FAM (Fluorescein, tagged with Y-shape DNA) and AuNPs. Based on the combination of FRET and signals circle amplification, the gold nanospheres functionalized with Y-motif DNA (Y@AuNPs) nanoprobe was utilized for effective ketamine detection with the limit of detection (LOD) down to 3 pg mL?1, which was lower than previously reported. Furthermore, the high sensitivity of Y@AuNPs facilitated quantitative analysis in biological media and practical samples.
关键词： Ultrasensitive detection,Ketamine,FRET,ATP-fueled target recycling,Fluorescent nanoprobe
更新于2025-09-16 10:30:52