A competitive immunoassay for detecting triazophos based on fluorescent catalytic hairpin self-assembly

Wang, Yuanshang; El‐Aty, A.M. Abd; Ge, Chunhua; Jia, Huiyan; Cui, Xueyan; Xu, Lingyuan; Cao, Zhen; She, Yongxin; Jin, Fen; Zhang, Yudan; Hacımüftüoğlu, Ahmet; Lamu, Sangqiong; Wang, Jing; Zheng, Lirong; Jin, Maojun; Hammock, Bruce D.

doi:10.1007/s00604-022-05217-5

Cited by 11 publications

(3 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…[25][26][27] In particular, catalytic hairpin self-assembly (CHA) is favored for its excellent specificity and sensitivity as a method to extend and amplify nucleotide sequences at room temperature. 28,29 In addition, enzyme-assisted signal amplification (EASA) uses polymerase and endonuclease as raw materials to exponentially amplify detected objects by the principle of semiconserved replication of DNA, resulting in signal amplification. 30,31 Although these methods can achieve signal amplification independently, the single-signal amplification strategy has a weak signal and low sensitivity in detecting some low-abundance substances.…”

Section: Introductionmentioning

confidence: 99%

Combined SERS Microfluidic Chip with Gold Nanocone Array for Effective Early Lung Cancer Prognosis in Mice Model

Qian¹,

Gu²,

Deng³

et al. 2023

IJN

View full text Add to dashboard Cite

Introduction:As the most common malignant tumor in the world, the prognosis of patients with advanced lung cancer remains poor even after treatment. There are many prognostic marker assays available, but there is still more room for the development of highthroughput and sensitive detection of circulating tumor DNA (ctDNA). Surface-enhanced Raman spectroscopy (SERS), a spectroscopic detection method that has received wide attention in recent years, can achieve exponential amplification of Raman signals by using different metallic nanomaterials. Integrating SERS with signal amplification strategy into the microfluidic chip and applying it to ctDNA detection is expected to be an effective tool for the prognosis of lung cancer treatment effect in the future. Methods: To construct a high-throughput SERS microfluidic chip integrated with enzyme-assisted signal amplification (EASA) and catalytic hairpin self-assembly (CHA) signal amplification strategies, using hpDNA-functionalized Au nanocone arrays (AuNCAs) as capture substrates and cisplatin-treated lung cancer mice to simulate the detection environment for sensitive detection of ctDNA in serum of lung cancer patients after treatment. Results:The SERS microfluidic chip constructed by this scheme, with two reaction zones, can simultaneously and sensitively detect the concentrations of four prognostic ctDNAs in the serum of three lung cancer patients with a limit of detection (LOD) as low as the aM level. The results of the ELISA assay are consistent with this scheme, and its accuracy is guaranteed. Conclusion: This high-throughput SERS microfluidic chip has high sensitivity and specificity in the detection of ctDNA. This could be a potential tool for prognostic assessment of lung cancer treatment efficacy in future clinical applications.

show abstract

Section: Introductionmentioning

confidence: 99%

Combined SERS Microfluidic Chip with Gold Nanocone Array for Effective Early Lung Cancer Prognosis in Mice Model

Qian¹,

Gu²,

Deng³

et al. 2023

IJN

View full text Add to dashboard Cite

show abstract

“…In recent years, metal NPs [Au, Ag, and Cu NPs] have been reported for their excellent performance as fluorescent probes in many assays because of their simple synthesis, excellent optical properties, good biocompatibility, and ultra-fine size. − Compared with AuNPs and AgNPs, Cu NPs are promising candidates for sensor construction, given their safety and the easily available copper sources . Momeni et al synthesized fluorescent Cu NPs in the presence of ascorbic acid with a small size and an average diameter of 10 nm and Cu NPs were successfully used to detect CN – in water samples .…”

Section: Introductionmentioning

confidence: 99%

Multifunctional Probe Based on CTAB-Cu Nanoparticles for Fluorescence and Colorimetric Dual-Read-Out Determination of p-Nitrophenol and Glyphosate

Wang

Peng

et al. 2023

ACS Sustainable Chem. Eng.

View full text Add to dashboard Cite

The quantitative analysis of glyphosate has significant implications for the evaluation of food quality and environmental safety risks. First, copper nanoparticles (Cu NPs) were synthesized using cetyltrimethylammonium bromide (CTAB) as a stabilizing agent, which resolved the limitation associated with atmospheric oxidation of Cu NPs and provided excellent fluorescence stability. Interestingly, the fluorescence of blue-emitting CTAB-Cu NPs was quenched by pH-responsive p-nitrophenol (p-NP) via Forster resonance energy transfer. Herein, a dual read-out sensor based on CTAB-Cu NPs and p-NP was constructed for glyphosate detection. Specifically, acetic acid (HAc) from acetylcholine (ACh) via hydrolysis catalyzed by acetyl cholinesterase (AChE) reduced the pH of the mixed solution and prevented the loss of protons from p-NP, which allowed the fluorescence emission by CTAB-Cu NPs. The presence of glyphosate inhibited the activity of AChE, and the increased pH resulted in proton loss. The fluorescence intensity of CTAB-Cu NPs was quenched, and the solution color changed from colorless to yellow with a UV absorption peak at 400 nm. The sensor can detect ultra-trace levels of glyphosate residues with a limit of detection of 5.43 ng/L. The obtained biosensor was successfully utilized to monitor the degradation of glyphosate in the roots of cabbage and cultivated soil.

show abstract

“…Thus, detection methods with higher sensitivity need to be developed. On this basis, some isothermal signal amplification strategies have been proposed, including the hybridization chain reaction [15], catalytic hairpin self-assembly (CHA) [16], and rolling circle amplification [17,18]. Despite their high application efficiency, harsh thermal annealing, expensive protocols and unnecessary by-products hinder their widespread application.…”

Section: Introductionmentioning

confidence: 99%

Self-Supplying Guide RNA-Mediated CRISPR/Cas12a Fluorescence System for Sensitive Detection of T4 PNKP

Yuan

Liu

et al. 2022

Molecules

View full text Add to dashboard Cite

Sensitive detection methods for T4 polynucleotide kinase/phosphatase (T4 PNKPP) are urgently required to obtain information on malignancy and thereby to provide better guidance in PNKP-related diagnostics and drug screening. Although the CRISPR/Cas12a system shows great promise in DNA-based signal amplification protocols, its guide RNAs with small molecular weight often suffer nuclease degradation during storage and utilization, resulting in reduced activation efficiency. Herein, we proposed a self-supplying guide RNA-mediated CRISPR/Cas12a system for the sensitive detection of T4 PNKP in cancer cells, in which multiple copies of guide RNA were generated by in situ transcription. In this assay, T4 PNKP was chosen as a model, and a dsDNA probe with T7 promoter region and the transcription region of guide RNA were involved. Under the action of T4 PNKP, the 5′-hydroxyl group of the dsDNA probe was converted to a phosphate group, which can be recognized and digested by Lambda Exo, resulting in dsDNA hydrolysis. The transcription template was destroyed, which resulted in the failure to generate guide RNA by the transcription pathway. Therefore, the CRISPR/Cas12a system could not be activated to effectively cleavage the F-Q-reporter, and the fluorescence signal was turned off. In the absence of T4 PNKP, the 5′-hydroxyl group of the substrate DNA cannot be digested by Lambda Exo. The intact dsDNA acts as the transcription template to generate a large amount of guide RNA. Finally, the formed Cas12a/gRNA complex triggered the reverse cleavage of Cas12a on the F-Q-reporter, resulting in a “turn-on” fluorescence signal. This strategy displayed sharp sensitivity of T4 PNKP with the limit of detection (LOD) down to 0.0017 mU/mL, which was mainly due to the multiple regulation effect of transcription amplification. In our system, the dsDNA simultaneously serves as the T4 PNKP substrate, transcription template, and Lambda Exo substrate, avoiding the need for multiple probe designs and saving costs. By integrating the target recognition, Lambda Exo activity, and trans-cleavage activity of Cas12a, CRISPR/Cas12a catalyzed the cleavage of fluorescent-labeled short-stranded DNA probes and enabled synergetic signal amplification for sensitive T4 PNKP detection. Furthermore, the T4 PNKP in cancer cells has been evaluated as a powerful tool for biomedical research and clinical diagnosis, proving a good practical application capacity.

show abstract

A competitive immunoassay for detecting triazophos based on fluorescent catalytic hairpin self-assembly

Cited by 11 publications

References 31 publications

Combined SERS Microfluidic Chip with Gold Nanocone Array for Effective Early Lung Cancer Prognosis in Mice Model

Combined SERS Microfluidic Chip with Gold Nanocone Array for Effective Early Lung Cancer Prognosis in Mice Model

Multifunctional Probe Based on CTAB-Cu Nanoparticles for Fluorescence and Colorimetric Dual-Read-Out Determination of p-Nitrophenol and Glyphosate

Self-Supplying Guide RNA-Mediated CRISPR/Cas12a Fluorescence System for Sensitive Detection of T4 PNKP

Contact Info

Product

Resources

About