Cascade CRISPR/cas enables amplification-free microRNA sensing with fM-sensitivity and single-base-specificity

2021 ◽  
Vol 57 (2) ◽  
pp. 247-250
Author(s):  
Yong Sha ◽  
Ru Huang ◽  
Mengqi Huang ◽  
Huahua Yue ◽  
Yuanyue Shan ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Diagnostic System ◽  
Single Base ◽  
Base Specificity ◽  
Mirna Detection

A cascade CRISPR/cas nucleic acid diagnostic system, which can achieve high-sensitive and single-base specificity without target amplification, was developed for miRNA detection.

scholarly journals A CRISPR-Cas autocatalysis-driven feedback amplification network for supersensitive DNA diagnostics

2021 ◽  
Vol 7 (5) ◽  
pp. eabc7802
Author(s):  
Kai Shi ◽  
Shiyi Xie ◽  
Renyun Tian ◽  
Shuo Wang ◽  
Qin Lu ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Genomic Dna ◽  
Reaction Network ◽  
Clinical Samples ◽  
Great Promise ◽  
Base Specificity ◽  
Nucleotide Mutation ◽  
Positive Feedback Circuit ◽  
Stringent Target ◽  
And Function

Artificial nucleic acid circuits with precisely controllable dynamic and function have shown great promise in biosensing, but their utility in molecular diagnostics is still restrained by the inability to process genomic DNA directly and moderate sensitivity. To address this limitation, we present a CRISPR-Cas–powered catalytic nucleic acid circuit, namely, CRISPR-Cas–only amplification network (CONAN), for isothermally amplified detection of genomic DNA. By integrating the stringent target recognition, helicase activity, and trans-cleavage activity of Cas12a, a Cas12a autocatalysis-driven artificial reaction network is programmed to construct a positive feedback circuit with exponential dynamic in CONAN. Consequently, CONAN achieves one-enzyme, one-step, real-time detection of genomic DNA with attomolar sensitivity. Moreover, CONAN increases the intrinsic single-base specificity of Cas12a, and enables the effective detection of hepatitis B virus infection and human bladder cancer–associated single-nucleotide mutation in clinical samples, highlighting its potential as a powerful tool for disease diagnostics.

A rapid, instrument-free, sample-to-result nucleic acid amplification test

Lab on a Chip ◽  
2016 ◽  
Vol 16 (19) ◽  
pp. 3777-3787 ◽  
Author(s):  
Lisa K. Lafleur ◽  
Joshua D. Bishop ◽  
Erin K. Heiniger ◽  
Ryan P. Gallagher ◽  
Maxwell D. Wheeler ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Diagnostic System ◽  
Nucleic Acid Amplification Test ◽  
Nucleic Acid Amplification ◽  
Test Device ◽  
Fully Integrated ◽  
Free Sample ◽  
System Prototype

This diagnostic system prototype is the first demonstration of a fully integrated, simple-to-use nucleic acid amplification test device.

scholarly journals Peptide nucleic acid-templated selenocystine–selenoester ligation enables rapid miRNA detection

2018 ◽  
Vol 9 (4) ◽  
pp. 896-903 ◽  
Author(s):  
Jessica Sayers ◽  
Richard J. Payne ◽  
Nicolas Winssinger
Keyword(s):  
Nucleic Acid ◽  
Peptide Nucleic Acid ◽  
Specific Detection ◽  
Ligation Reaction ◽  
Low Concentrations ◽  
Mirna Detection ◽  
Peptide Ligation

A PNA-templated peptide ligation reaction has been developed between selenocystine and selenoesters. The methodology was used for the sequence specific detection of miRNA at low concentrations.

A biosensor for the detection of single base mismatches in microRNA

2015 ◽  
Vol 51 (78) ◽  
pp. 14597-14600 ◽  
Author(s):  
Jieon Lee ◽  
Ginam Park ◽  
Dal-Hee Min
Keyword(s):  
Graphene Oxide ◽  
Nucleic Acid ◽  
Nucleic Acid Detection ◽  
Single Base ◽  
Target Probe

Graphene oxide enables highly sequence specific nucleic acid detection by selectively removing the signal from a mismatched target/probe duplex.

scholarly journals 3′ Tth Endonuclease Cleavage Polymerase Chain Reaction (3TEC-PCR) Technology for Single-Base-Specific Multiplex Pathogen Detection using a Two-Oligonucleotide System

2021 ◽  
Vol 22 (11) ◽  
pp. 6061
Author(s):  
Owen Higgins ◽  
Terry J. Smith
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Target Detection ◽  
Pathogen Detection ◽  
Multiplex Detection ◽  
Chain Reaction ◽  
Single Base ◽  
Base Specificity ◽  
Pcr Technology ◽  
Polymerase Chain

Polymerase chain reaction (PCR) is the standard in nucleic acid amplification technology for infectious disease pathogen detection and has been the primary diagnostic tool employed during the global COVID-19 pandemic. Various PCR technology adaptations, typically using two-oligonucleotide dye-binding methods or three-oligonucleotide hydrolysis probe systems, enable real-time multiplex target detection or single-base specificity for the identification of single-nucleotide polymorphisms (SNPs). A small number of two-oligonucleotide PCR systems facilitating both multiplex detection and SNP identification have been reported; however, these methods often have limitations in terms of target specificity, production of variable or false-positive results, and the requirement for extensive optimisation or post-amplification analysis. This study introduces 3′ Tth endonuclease cleavage PCR (3TEC-PCR), a two-oligonucleotide PCR system incorporating a modified primer/probe and a thermostable cleavage enzyme, Tth endonuclease IV, for real-time multiplex detection and SNP identification. Complete analytical specificity, low limits of detection, single-base specificity, and simultaneous multiple target detection have been demonstrated in this study using 3TEC-PCR to identify bacterial meningitis associated pathogens. This is the first report of a two-oligonucleotide, real-time multiplex PCR technology with single-base specificity using Tth endonuclease IV.

Electrical Detection of DNA Hybridization with Single-Base Specificity Using Transistors Based on CVD-Grown Graphene Sheets

2010 ◽  
Vol 22 (14) ◽  
pp. 1649-1653 ◽  
Author(s):  
Xiaochen Dong ◽  
Yumeng Shi ◽  
Wei Huang ◽  
Peng Chen ◽  
Lain-Jong Li
Keyword(s):  
Dna Hybridization ◽  
Graphene Sheets ◽  
Electrical Detection ◽  
Single Base ◽  
Base Specificity ◽  
Grown Graphene

scholarly journals Polymerase-free measurement of microRNA-122 with single base specificity using single molecule arrays: Detection of drug-induced liver injury

PLoS ONE ◽  
2017 ◽  
Vol 12 (7) ◽  
pp. e0179669 ◽  
Author(s):  
David M. Rissin ◽  
Barbara López-Longarela ◽  
Salvatore Pernagallo ◽  
Hugh Ilyine ◽  
A. D. Bastiaan Vliegenthart ◽  
...  
Keyword(s):  
Liver Injury ◽  
Single Molecule ◽  
Drug Induced ◽  
Single Base ◽  
Drug Induced Liver Injury ◽  
Base Specificity
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