Isothermal Detection of DNA by Beacon‐Assisted Detection Amplification

Abstract

Ein einfacher Biochip aus zwei integrierten molekularen Schaltern ermöglicht den Nachweis und die Vervielfältigung spezifischer Nucleinsäuren in biologischen Extrakten. Die Nucleinsäure wird mithilfe eines Beacons detektiert, anschließend werden eine DNA‐Polymerase und Nicking‐Endonuclease zur schnellen Signalverstärkung für den empfindlichen Nucleinsäurenachweis verwendet.

Ratiometric fluorescence transduction by hybridization after isothermal amplification for determination of zeptomole quantities of oligonucleotide biomarkers with a paper-based platform and camera-based detection

“…Examples of isothermal methods that have been reported for amplification of nucleic acid targets are rolling circle amplification [15], loop-mediated amplification [16,17], helicase dependent amplification [18,19], exponential amplification reaction [20,21] and exonuclease-catalyzed target recycling [22,23]. Amplification of target concentration using isothermal methods also facilitates an integration of a readout platform that is of lower cost and technical complexity, making the diagnostic platform more appealing for implementation in resource-limited settings and for point-of-care diagnosis.…”

Circular Strand Displacement Polymerization Reaction: A Promising Technique?

“…Some researchers have tried to introduce nicking endonuclease with two molecular switches operating in series in order to drive an exponential amplification [8]. This homogeneous assay includes an amplification element, a detection transducer and a dye of acceptable specificity, as well as a format that eliminates the need for opening tube lids to avoid post-amplification manipulation and contamination.…”

Cleaved DNAzyme substrate induced enzymatic cascade for the exponential amplified analysis of l-histidine

“…In the nucleic acid-based enzymatic cascade assays, the exponential isothermal amplification of DNA often employs a strand displacement polymerase and a single-strand nicking enzyme to continuously replicate one strand of a DNA duplex under a constant temperature [4][5][6][7][8]. Since the target can activate circular polymerization reactions, enzymatic cascade mediated strategy offers a versatile method with high sensitivity and specificity.…”