Cascade CRISPR/cas enables amplification-free microRNA sensing with fM-sensitivity and single-base-specificity
نویسندگان
چکیده
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.
منابع مشابه
Polymerase-free measurement of microRNA-122 with single base specificity using single molecule arrays: Detection of drug-induced liver injury
We have developed a single probe method for detecting microRNA from human serum using single molecule arrays, with sequence specificity down to a single base, and without the use of amplification by polymerases. An abasic peptide nucleic acid (PNA) probe-containing a reactive amine instead of a nucleotide at a specific position in the sequence-for detecting a microRNA was conjugated to superpar...
متن کاملSensitivity and Specificity of Nucleic Acid Sequence-Based Amplification Method for Diagnosis of Cutaneous Leishmaniasis
Abstract Background and Objective: Culture, microscopic method is a gold standard method for identification of Lishmania parasite. The use of Molecular methods such as RT- PCR compared to microscopic methods has a higher sensitivity and specificity however, it is not widely used due to its expensive equipment and the time requested. The use of nucleic acid sequence based amplification (NASBA) ...
متن کاملTarget-triggered cascade recycling amplification for label-free detection of microRNA and molecular logic operations.
A cascade recycling amplification (CRA) that implements cascade logic circuits with feedback amplification function is developed for label-free chemiluminescence detection of microRNA-122 with an ultrahigh sensitivity of 0.82 fM and excellent specificity, which is applied to construct a series of molecular-scale two-input logic gates by using microRNAs as inputs and CRA products as outputs.
متن کاملSensitive and homogeneous microRNA detection using branched cascade enzymatic amplification.
This assay, termed branched cascade enzymatic amplification (BCEA), can be a novel and straightforward method for sensitive and specific microRNA detection in crude cellular extracts of cancer cells at physiological temperature, by coupling two ordinary polymerases, Klenow fragment exo(-) and terminal deoxynucleotidyl transferase.
متن کاملMaximizing sensitivity and specificity of PCR by pre-amplification heating
We have found that assembling the reaction mixture at a temperature greater than the annealing temperature improved both product yield and specificity of PCR. When reactions were maintained at 70°C in a dry heating block during addition of denatured samples to aliquotted reagent master mix, a reproducible increase in product yield was observed compared to duplicates maintained at room temperatu...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Chemical Communications
سال: 2021
ISSN: ['1359-7345', '1364-548X']
DOI: https://doi.org/10.1039/d0cc06412b