scholarly journals A nuclease protection ELISA assay for colorimetric and electrochemical detection of nucleic acids

2019 ◽  
Vol 11 (8) ◽  
pp. 1027-1034 ◽  
Author(s):  
Jessica E. Filer ◽  
Robert B. Channon ◽  
Charles S. Henry ◽  
Brian J. Geiss

The NP-ELISA combines traditional nuclease protection with optical and electrochemical enzymatic readout for nucleic acid detection.

2021 ◽  
Author(s):  
hongyu liu ◽  
Yuhao You ◽  
Youzhuo Zhu ◽  
Heng Zheng

Detection of nucleic acids have become significantly important in molecular diagnostics, genetics therapy, mutation analysis, forensic investigations and biomedical development, and so on. In recent years, exonuclease Ⅲ (Exo III)...


The Analyst ◽  
2019 ◽  
Vol 144 (20) ◽  
pp. 5923-5927 ◽  
Author(s):  
Shuang Liu ◽  
Chen Xin ◽  
Xiaoxiao Yu ◽  
Zhenbo Ding ◽  
Shufeng Liu

A catalytic DNA circuit-programmed and enzyme-powered autonomous DNA machine was proposed for one-step, isothermal and dual-level amplified detection of nucleic acids.


Lab on a Chip ◽  
2015 ◽  
Vol 15 (7) ◽  
pp. 1697-1707 ◽  
Author(s):  
Mark D. Borysiak ◽  
Kevin W. Kimura ◽  
Jonathan D. Posner

The NAIL device integrates isotachophoresis and loop-mediated isothermal amplification (LAMP) with mobile phone detection to extract, amplify, and detect nucleic acids from complex matrices in less than one hour.


2015 ◽  
Author(s):  
◽  
Kai Tian

[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT REQUEST OF AUTHOR.] The nanopore sensor can detect cancer-derived nucleic acid biomarkers such as microRNAs (miRNAs), providing a noninvasive tool potentially useful in medical diagnostics. However, the nanopore-based detection of these biomarkers remains confounded by the presence of numerous other nucleic acid species found in biofluid extracts. Their nonspecific interactions with the nanopore inevitably contaminate the target signals, reducing the detection accuracy. Here we report a novel method that utilizes a polycationic peptide-PNA probe as the carrier for selective nucleic acid detection in the nucleic acids mixture. The cationic probe hybridized with DNA or RNA forms a dipole complex, which can be captured by the pore using a voltage polarity that is opposite the polarity used to capture negatively charged nucleic acids. As a result, non-target species are driven away from the pore opening, and the target sequences can be detected accurately without interference. In addition, we demonstrate that the PNA probe enables to accurately discriminate single-nucleotide difference. Moreover, molecule dynamic simulation is applied to expose the mechanism. Combined with experimental and calculating data, we construct a model to demonstrate that it is universal for all kinds of nucleic acid targets. In sum, this highly sensitive and selective nano-dielectrophoresis approach can be applied to the detection of clinically relevant nucleic acid fragments in complex samples and fulfills the diagnostic of diseases in early stage.


2009 ◽  
Vol 55 (4) ◽  
pp. 670-683 ◽  
Author(s):  
Larry J Kricka ◽  
Paolo Fortina

Abstract Background: The inherent fluorescent properties of nucleosides, nucleotides, and nucleic acids are limited, and thus the need has arisen for fluorescent labeling of these molecules for a variety of analytical applications. Content: This review traces the analytical ancestry of fluorescent labeling of nucleosides, nucleotides, and nucleic acids, with an emphasis on the first to publish or patent. The scope of labeling includes (a) direct labeling by covalent labeling of nucleic acids with a fluorescent label or noncovalent binding or intercalation of a fluorescent dye to nucleic acids and (b) indirect labeling via covalent attachment of a secondary label to a nucleic acid, and then binding this to a fluorescently labeled ligand binder. An alternative indirect strategy involves binding of a nucleic acid to a nucleic acid binder molecule (e.g., antibody, antibiotic, histone, antibody, nuclease) that is labeled with a fluorophore. Fluorescent labels for nucleic acids include organic fluorescent dyes, metal chelates, carbon nanotubes, quantum dots, gold particles, and fluorescent minerals. Summary: Fluorescently labeled nucleosides, nucleotides, and nucleic acids are important types of reagents for biological assay methods and underpin current methods of chromosome analysis, gel staining, DNA sequencing and quantitative PCR. Although these methods use predominantly organic fluorophores, new types of particulate fluorophores in the form of nanoparticles, nanorods, and nanotubes may provide the basis of a new generation of fluorescent labels and nucleic acid detection methods.


2019 ◽  
Vol 16 (3) ◽  
pp. 235-239
Author(s):  
Masood Ayoub ◽  
Bilal Ahmad Bhat ◽  
Shahjahan Ul Islam ◽  
Syed Masood Ahmad Rizvi ◽  
Qazi Mohd Junaid

The design and development of synthetic fluorescent molecular architectures for sensing of nucleic acids and related species in living cells is an area of enormous interest. For the first time a novel compilation of single molecular abiotic fluorescent receptors for nucleic acid detection in living cells have been reviewed. Selected reports have been screened and thoroughly discussed which have revealed enormous promise for bio imaging. The mechanistic aspects of nucleic acid, phosphate or nitrogenous base sensing upon encounter with the receptors has been examined under diverse matrices. In addition to the cytotoxicity, specific conditions deciphering suitable and promising results for real-time application have been highlighted.


Author(s):  
Tang Guanghui ◽  
Jiaojiao Gong ◽  
Lijuan Kan ◽  
Xiuming Zhang ◽  
Ying He ◽  
...  

Abstract CRISPR/Cas12a system has been shown promising for nucleic acid diagnostics due to its rapid, portable and accurate features. In combination with isothermal amplification technology, single-copy sensitivity can be achieved. However, cleavage of the amplicons and primers by the cis- and trans-activity of Cas12a hinders the attempts to integrate the amplification and detection steps into a single reaction. Through phosphorothioate modification of primer and design of crRNA that allow for the cutting site locating at the modified site of the primer, we realized onepot detection of SARS-CoV-2 with single-copy sensitivity. We also identified the activated Cas12a has a much higher affinity to C nucleotide-rich reporter than others. By applying such reporters, we significantly reduced the reaction time required for the lateral-flow readout. Furthermore, to improve the specificity of the strip-based assay, we created a novel reporter and, when combined with a customized strip, the unspecific signal could be completely eliminated. This established system termed Targeting DNA by Cas12a-based Eye Sight Testing in Onepot Reaction (TESTOR) was validated using clinical cervical samples for human papillomaviruses (HPVs) detection. Our system represents a general approach to integrating the nucleic acid amplification and detection into a onepot reaction in CRISPR-Cas systems, highlighting its potential as a rapid, portable and accurate detection platform of nucleic acids.


The Analyst ◽  
2018 ◽  
Vol 143 (19) ◽  
pp. 4559-4565 ◽  
Author(s):  
Junjie Chi ◽  
Biao Ma ◽  
Xing Dong ◽  
Bingbing Gao ◽  
Abdelrahman Elbaz ◽  
...  

Here we report a bio-inspired photonic nitrocellulose array for ultrasensitive nucleic-acid detection.


The Analyst ◽  
2015 ◽  
Vol 140 (13) ◽  
pp. 4370-4373 ◽  
Author(s):  
Xiaodong Ma ◽  
Weiwei Xu ◽  
Chao Chen ◽  
Zuhong Lu ◽  
Jiong Li

A nanoliter droplet array based on a hydrophilic–hydrophobic patterned chip is developed without using microfabrication technology, which could be applied to detect nucleic acids.


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